PubMed:15190003 JSONTXT

{"project":"Glycan-Motif","denotations":[{"id":"T1","span":{"begin":182,"end":191},"obj":"https://glytoucan.org/Structures/Glycans/G65889KE"},{"id":"T2","span":{"begin":182,"end":191},"obj":"https://glytoucan.org/Structures/Glycans/G68158BT"},{"id":"T3","span":{"begin":1276,"end":1283},"obj":"https://glytoucan.org/Structures/Glycans/G15021LG"},{"id":"T4","span":{"begin":1422,"end":1431},"obj":"https://glytoucan.org/Structures/Glycans/G65889KE"},{"id":"T5","span":{"begin":1422,"end":1431},"obj":"https://glytoucan.org/Structures/Glycans/G68158BT"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"GlyCosmos6-Glycan-Motif-Image","denotations":[{"id":"T1","span":{"begin":182,"end":191},"obj":"Glycan_Motif"},{"id":"T3","span":{"begin":1276,"end":1283},"obj":"Glycan_Motif"},{"id":"T4","span":{"begin":1422,"end":1431},"obj":"Glycan_Motif"}],"attributes":[{"id":"A1","pred":"image","subj":"T1","obj":"https://api.glycosmos.org/wurcs2image/0.10.0/png/binary/G68158BT"},{"id":"A2","pred":"image","subj":"T1","obj":"https://api.glycosmos.org/wurcs2image/0.10.0/png/binary/G65889KE"},{"id":"A3","pred":"image","subj":"T3","obj":"https://api.glycosmos.org/wurcs2image/0.10.0/png/binary/G15021LG"},{"id":"A4","pred":"image","subj":"T4","obj":"https://api.glycosmos.org/wurcs2image/0.10.0/png/binary/G68158BT"},{"id":"A5","pred":"image","subj":"T4","obj":"https://api.glycosmos.org/wurcs2image/0.10.0/png/binary/G65889KE"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"sentences","denotations":[{"id":"TextSentencer_T1","span":{"begin":0,"end":192},"obj":"Sentence"},{"id":"TextSentencer_T2","span":{"begin":193,"end":350},"obj":"Sentence"},{"id":"TextSentencer_T3","span":{"begin":351,"end":534},"obj":"Sentence"},{"id":"TextSentencer_T4","span":{"begin":535,"end":664},"obj":"Sentence"},{"id":"TextSentencer_T5","span":{"begin":665,"end":799},"obj":"Sentence"},{"id":"TextSentencer_T6","span":{"begin":800,"end":989},"obj":"Sentence"},{"id":"TextSentencer_T7","span":{"begin":990,"end":1192},"obj":"Sentence"},{"id":"TextSentencer_T8","span":{"begin":1193,"end":1441},"obj":"Sentence"},{"id":"TextSentencer_T9","span":{"begin":1442,"end":1710},"obj":"Sentence"},{"id":"T1","span":{"begin":0,"end":192},"obj":"Sentence"},{"id":"T2","span":{"begin":193,"end":350},"obj":"Sentence"},{"id":"T3","span":{"begin":351,"end":534},"obj":"Sentence"},{"id":"T4","span":{"begin":535,"end":664},"obj":"Sentence"},{"id":"T5","span":{"begin":665,"end":799},"obj":"Sentence"},{"id":"T6","span":{"begin":800,"end":989},"obj":"Sentence"},{"id":"T7","span":{"begin":990,"end":1192},"obj":"Sentence"},{"id":"T8","span":{"begin":1193,"end":1441},"obj":"Sentence"},{"id":"T9","span":{"begin":1442,"end":1710},"obj":"Sentence"},{"id":"T1","span":{"begin":0,"end":192},"obj":"Sentence"},{"id":"T2","span":{"begin":193,"end":350},"obj":"Sentence"},{"id":"T3","span":{"begin":351,"end":534},"obj":"Sentence"},{"id":"T4","span":{"begin":535,"end":664},"obj":"Sentence"},{"id":"T5","span":{"begin":665,"end":799},"obj":"Sentence"},{"id":"T6","span":{"begin":800,"end":989},"obj":"Sentence"},{"id":"T7","span":{"begin":990,"end":1192},"obj":"Sentence"},{"id":"T8","span":{"begin":1193,"end":1441},"obj":"Sentence"},{"id":"T9","span":{"begin":1442,"end":1710},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"GlyCosmos6-Glycan-Motif-Structure","denotations":[{"id":"T1","span":{"begin":182,"end":191},"obj":"https://glytoucan.org/Structures/Glycans/G65889KE"},{"id":"T2","span":{"begin":182,"end":191},"obj":"https://glytoucan.org/Structures/Glycans/G68158BT"},{"id":"T3","span":{"begin":1276,"end":1283},"obj":"https://glytoucan.org/Structures/Glycans/G15021LG"},{"id":"T4","span":{"begin":1422,"end":1431},"obj":"https://glytoucan.org/Structures/Glycans/G65889KE"},{"id":"T5","span":{"begin":1422,"end":1431},"obj":"https://glytoucan.org/Structures/Glycans/G68158BT"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"uniprot-human","denotations":[{"id":"T1","span":{"begin":851,"end":853},"obj":"http://www.uniprot.org/uniprot/Q15181"},{"id":"T2","span":{"begin":851,"end":853},"obj":"http://www.uniprot.org/uniprot/P01298"},{"id":"T3","span":{"begin":1272,"end":1295},"obj":"http://www.uniprot.org/uniprot/Q14376"},{"id":"T4","span":{"begin":1320,"end":1331},"obj":"http://www.uniprot.org/uniprot/Q99484"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"uniprot-mouse","denotations":[{"id":"T1","span":{"begin":435,"end":441},"obj":"http://www.uniprot.org/uniprot/Q9ESF4"},{"id":"T2","span":{"begin":435,"end":441},"obj":"http://www.uniprot.org/uniprot/P54227"},{"id":"T3","span":{"begin":851,"end":853},"obj":"http://www.uniprot.org/uniprot/P10601"},{"id":"T4","span":{"begin":1272,"end":1295},"obj":"http://www.uniprot.org/uniprot/Q8R059"},{"id":"T5","span":{"begin":1320,"end":1331},"obj":"http://www.uniprot.org/uniprot/P38649"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"GlycoBiology-NCBITAXON","denotations":[{"id":"T1","span":{"begin":105,"end":111},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/1365831"},{"id":"T2","span":{"begin":221,"end":231},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/2759"},{"id":"T3","span":{"begin":330,"end":349},"obj":"http://purl.bioontology.org/ontology/STY/T039"},{"id":"T4","span":{"begin":435,"end":441},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/254545"},{"id":"T5","span":{"begin":738,"end":744},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/1365831"},{"id":"T6","span":{"begin":1300,"end":1304},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/158455"},{"id":"T7","span":{"begin":1300,"end":1304},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/3554"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"GO-BP","denotations":[{"id":"T1","span":{"begin":29,"end":42},"obj":"http://purl.obolibrary.org/obo/GO_0070085"},{"id":"T2","span":{"begin":292,"end":305},"obj":"http://purl.obolibrary.org/obo/GO_0070085"},{"id":"T3","span":{"begin":489,"end":502},"obj":"http://purl.obolibrary.org/obo/GO_0070085"},{"id":"T4","span":{"begin":614,"end":627},"obj":"http://purl.obolibrary.org/obo/GO_0070085"},{"id":"T5","span":{"begin":713,"end":726},"obj":"http://purl.obolibrary.org/obo/GO_0070085"},{"id":"T6","span":{"begin":249,"end":280},"obj":"http://purl.obolibrary.org/obo/GO_0043687"},{"id":"T7","span":{"begin":563,"end":571},"obj":"http://purl.obolibrary.org/obo/GO_0007349"},{"id":"T8","span":{"begin":1230,"end":1245},"obj":"http://purl.obolibrary.org/obo/GO_0051645"},{"id":"T9","span":{"begin":1236,"end":1245},"obj":"http://purl.obolibrary.org/obo/GO_0051179"},{"id":"T10","span":{"begin":1320,"end":1342},"obj":"http://purl.obolibrary.org/obo/GO_0004380"},{"id":"T11","span":{"begin":1320,"end":1342},"obj":"http://purl.obolibrary.org/obo/GO_0016740"},{"id":"T12","span":{"begin":1320,"end":1342},"obj":"http://purl.obolibrary.org/obo/GO_0051347"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"GO-CC","denotations":[{"id":"T1","span":{"begin":62,"end":66},"obj":"http://purl.obolibrary.org/obo/GO_0019013"},{"id":"T2","span":{"begin":1230,"end":1235},"obj":"http://purl.obolibrary.org/obo/GO_0005794"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"NGLY1-deficiency","denotations":[{"id":"PD-NGLY1-deficiency-B_T1","span":{"begin":841,"end":847},"obj":"chem:24139"},{"id":"PD-NGLY1-deficiency-B_T2","span":{"begin":963,"end":969},"obj":"chem:24139"},{"id":"PD-NGLY1-deficiency-B_T3","span":{"begin":1131,"end":1137},"obj":"chem:24139"},{"id":"PD-NGLY1-deficiency-B_T4","span":{"begin":1146,"end":1152},"obj":"chem:24139"}],"namespaces":[{"prefix":"hgnc","uri":"https://www.genenames.org/data/gene-symbol-report/#!/hgnc_id/HGNC:"},{"prefix":"omim","uri":"https://www.omim.org/entry/"},{"prefix":"chem","uri":"https://pubchem.ncbi.nlm.nih.gov/compound/"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"GlycoBiology-Motifs","denotations":[{"id":"T1","span":{"begin":421,"end":430},"obj":"http://rdf.glycoinfo.org/glycan/G00027MO"},{"id":"T2","span":{"begin":979,"end":988},"obj":"http://rdf.glycoinfo.org/glycan/G00027MO"},{"id":"T3","span":{"begin":1156,"end":1165},"obj":"http://rdf.glycoinfo.org/glycan/G00027MO"},{"id":"T4","span":{"begin":1700,"end":1709},"obj":"http://rdf.glycoinfo.org/glycan/G00027MO"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"Lectin","denotations":[{"id":"Lectin_T1","span":{"begin":1140,"end":1143},"obj":"https://acgg.asia/db/lfdb/LfDB0217"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"GlyTouCan-IUPAC","denotations":[{"id":"GlycanIUPAC_T1","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G26693XF\""},{"id":"GlycanIUPAC_T2","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G26693XF\""},{"id":"GlycanIUPAC_T3","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G26693XF\""},{"id":"GlycanIUPAC_T4","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G26693XF\""},{"id":"GlycanIUPAC_T5","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G01864SU\""},{"id":"GlycanIUPAC_T6","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G01864SU\""},{"id":"GlycanIUPAC_T7","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G01864SU\""},{"id":"GlycanIUPAC_T8","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G01864SU\""},{"id":"GlycanIUPAC_T9","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G17605FD\""},{"id":"GlycanIUPAC_T10","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G17605FD\""},{"id":"GlycanIUPAC_T11","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G17605FD\""},{"id":"GlycanIUPAC_T12","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G17605FD\""},{"id":"GlycanIUPAC_T13","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G41950LU\""},{"id":"GlycanIUPAC_T14","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G41950LU\""},{"id":"GlycanIUPAC_T15","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G41950LU\""},{"id":"GlycanIUPAC_T16","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G41950LU\""},{"id":"GlycanIUPAC_T17","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G57195RJ\""},{"id":"GlycanIUPAC_T18","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G57195RJ\""},{"id":"GlycanIUPAC_T19","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G57195RJ\""},{"id":"GlycanIUPAC_T20","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G57195RJ\""},{"id":"GlycanIUPAC_T21","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G85391SA\""},{"id":"GlycanIUPAC_T22","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G85391SA\""},{"id":"GlycanIUPAC_T23","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G85391SA\""},{"id":"GlycanIUPAC_T24","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G85391SA\""},{"id":"GlycanIUPAC_T25","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G89565QL\""},{"id":"GlycanIUPAC_T26","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G89565QL\""},{"id":"GlycanIUPAC_T27","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G89565QL\""},{"id":"GlycanIUPAC_T28","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G89565QL\""},{"id":"GlycanIUPAC_T29","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G80869MR\""},{"id":"GlycanIUPAC_T30","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G80869MR\""},{"id":"GlycanIUPAC_T31","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G80869MR\""},{"id":"GlycanIUPAC_T32","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G80869MR\""},{"id":"GlycanIUPAC_T33","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G55978NL\""},{"id":"GlycanIUPAC_T34","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G55978NL\""},{"id":"GlycanIUPAC_T35","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G55978NL\""},{"id":"GlycanIUPAC_T36","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G55978NL\""},{"id":"GlycanIUPAC_T37","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G54644LT\""},{"id":"GlycanIUPAC_T38","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G54644LT\""},{"id":"GlycanIUPAC_T39","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G54644LT\""},{"id":"GlycanIUPAC_T40","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G54644LT\""},{"id":"GlycanIUPAC_T41","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G25694UG\""},{"id":"GlycanIUPAC_T42","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G25694UG\""},{"id":"GlycanIUPAC_T43","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G25694UG\""},{"id":"GlycanIUPAC_T44","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G25694UG\""},{"id":"GlycanIUPAC_T45","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G25126RB\""},{"id":"GlycanIUPAC_T46","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G25126RB\""},{"id":"GlycanIUPAC_T47","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G25126RB\""},{"id":"GlycanIUPAC_T48","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G25126RB\""},{"id":"GlycanIUPAC_T49","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G51848AD\""},{"id":"GlycanIUPAC_T50","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G51848AD\""},{"id":"GlycanIUPAC_T51","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G51848AD\""},{"id":"GlycanIUPAC_T52","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G51848AD\""},{"id":"GlycanIUPAC_T53","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G94667GM\""},{"id":"GlycanIUPAC_T54","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G94667GM\""},{"id":"GlycanIUPAC_T55","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G94667GM\""},{"id":"GlycanIUPAC_T56","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G94667GM\""},{"id":"GlycanIUPAC_T57","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G30124BO\""},{"id":"GlycanIUPAC_T58","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G30124BO\""},{"id":"GlycanIUPAC_T59","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G30124BO\""},{"id":"GlycanIUPAC_T60","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G30124BO\""},{"id":"GlycanIUPAC_T61","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G82777EZ\""},{"id":"GlycanIUPAC_T62","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G82777EZ\""},{"id":"GlycanIUPAC_T63","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G82777EZ\""},{"id":"GlycanIUPAC_T64","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G82777EZ\""},{"id":"GlycanIUPAC_T65","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G10151YZ\""},{"id":"GlycanIUPAC_T66","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G10151YZ\""},{"id":"GlycanIUPAC_T67","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G10151YZ\""},{"id":"GlycanIUPAC_T68","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G10151YZ\""},{"id":"GlycanIUPAC_T69","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G17585ZM\""},{"id":"GlycanIUPAC_T70","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G17585ZM\""},{"id":"GlycanIUPAC_T71","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G17585ZM\""},{"id":"GlycanIUPAC_T72","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G17585ZM\""},{"id":"GlycanIUPAC_T73","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G04411CJ\""},{"id":"GlycanIUPAC_T74","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G04411CJ\""},{"id":"GlycanIUPAC_T75","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G04411CJ\""},{"id":"GlycanIUPAC_T76","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G04411CJ\""},{"id":"GlycanIUPAC_T77","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G38254HJ\""},{"id":"GlycanIUPAC_T78","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G38254HJ\""},{"id":"GlycanIUPAC_T79","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G38254HJ\""},{"id":"GlycanIUPAC_T80","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G38254HJ\""},{"id":"GlycanIUPAC_T81","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G75188FS\""},{"id":"GlycanIUPAC_T82","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G75188FS\""},{"id":"GlycanIUPAC_T83","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G75188FS\""},{"id":"GlycanIUPAC_T84","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G75188FS\""},{"id":"GlycanIUPAC_T85","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G70374VG\""},{"id":"GlycanIUPAC_T86","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G70374VG\""},{"id":"GlycanIUPAC_T87","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G70374VG\""},{"id":"GlycanIUPAC_T88","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G70374VG\""},{"id":"GlycanIUPAC_T89","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G45176LJ\""},{"id":"GlycanIUPAC_T90","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G45176LJ\""},{"id":"GlycanIUPAC_T91","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G45176LJ\""},{"id":"GlycanIUPAC_T92","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G45176LJ\""},{"id":"GlycanIUPAC_T93","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G30874VW\""},{"id":"GlycanIUPAC_T94","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G30874VW\""},{"id":"GlycanIUPAC_T95","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G30874VW\""},{"id":"GlycanIUPAC_T96","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G30874VW\""},{"id":"GlycanIUPAC_T97","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G69333MI\""},{"id":"GlycanIUPAC_T98","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G69333MI\""},{"id":"GlycanIUPAC_T99","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G69333MI\""},{"id":"GlycanIUPAC_T100","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G69333MI\""},{"id":"GlycanIUPAC_T101","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G10676XO\""},{"id":"GlycanIUPAC_T102","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G10676XO\""},{"id":"GlycanIUPAC_T103","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G10676XO\""},{"id":"GlycanIUPAC_T104","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G10676XO\""},{"id":"GlycanIUPAC_T105","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G14843DJ\""},{"id":"GlycanIUPAC_T106","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G14843DJ\""},{"id":"GlycanIUPAC_T107","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G14843DJ\""},{"id":"GlycanIUPAC_T108","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G14843DJ\""},{"id":"GlycanIUPAC_T109","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G47546FR\""},{"id":"GlycanIUPAC_T110","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G47546FR\""},{"id":"GlycanIUPAC_T111","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G47546FR\""},{"id":"GlycanIUPAC_T112","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G47546FR\""},{"id":"GlycanIUPAC_T113","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G73695ZM\""},{"id":"GlycanIUPAC_T114","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G73695ZM\""},{"id":"GlycanIUPAC_T115","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G73695ZM\""},{"id":"GlycanIUPAC_T116","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G73695ZM\""},{"id":"GlycanIUPAC_T117","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G31923TJ\""},{"id":"GlycanIUPAC_T118","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G31923TJ\""},{"id":"GlycanIUPAC_T119","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G31923TJ\""},{"id":"GlycanIUPAC_T120","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G31923TJ\""},{"id":"GlycanIUPAC_T121","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G60519EP\""},{"id":"GlycanIUPAC_T122","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G60519EP\""},{"id":"GlycanIUPAC_T123","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G60519EP\""},{"id":"GlycanIUPAC_T124","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G60519EP\""},{"id":"GlycanIUPAC_T125","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G07933IA\""},{"id":"GlycanIUPAC_T126","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G07933IA\""},{"id":"GlycanIUPAC_T127","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G07933IA\""},{"id":"GlycanIUPAC_T128","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G07933IA\""},{"id":"GlycanIUPAC_T129","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G40745NH\""},{"id":"GlycanIUPAC_T130","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G40745NH\""},{"id":"GlycanIUPAC_T131","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G40745NH\""},{"id":"GlycanIUPAC_T132","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G40745NH\""},{"id":"GlycanIUPAC_T133","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G54496YV\""},{"id":"GlycanIUPAC_T134","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G54496YV\""},{"id":"GlycanIUPAC_T135","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G54496YV\""},{"id":"GlycanIUPAC_T136","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G54496YV\""},{"id":"GlycanIUPAC_T137","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G62953SQ\""},{"id":"GlycanIUPAC_T138","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G62953SQ\""},{"id":"GlycanIUPAC_T139","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G62953SQ\""},{"id":"GlycanIUPAC_T140","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G62953SQ\""},{"id":"GlycanIUPAC_T141","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G70070AY\""},{"id":"GlycanIUPAC_T142","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G70070AY\""},{"id":"GlycanIUPAC_T143","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G70070AY\""},{"id":"GlycanIUPAC_T144","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G70070AY\""},{"id":"GlycanIUPAC_T145","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G78792WC\""},{"id":"GlycanIUPAC_T146","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G78792WC\""},{"id":"GlycanIUPAC_T147","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G78792WC\""},{"id":"GlycanIUPAC_T148","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G78792WC\""},{"id":"GlycanIUPAC_T149","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G25238AV\""},{"id":"GlycanIUPAC_T150","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G25238AV\""},{"id":"GlycanIUPAC_T151","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G25238AV\""},{"id":"GlycanIUPAC_T152","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G25238AV\""},{"id":"GlycanIUPAC_T153","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G40510DP\""},{"id":"GlycanIUPAC_T154","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G40510DP\""},{"id":"GlycanIUPAC_T155","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G40510DP\""},{"id":"GlycanIUPAC_T156","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G40510DP\""},{"id":"GlycanIUPAC_T157","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G61120TK\""},{"id":"GlycanIUPAC_T158","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G61120TK\""},{"id":"GlycanIUPAC_T159","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G61120TK\""},{"id":"GlycanIUPAC_T160","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G61120TK\""},{"id":"GlycanIUPAC_T161","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G41342KV\""},{"id":"GlycanIUPAC_T162","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G41342KV\""},{"id":"GlycanIUPAC_T163","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G41342KV\""},{"id":"GlycanIUPAC_T164","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G41342KV\""},{"id":"GlycanIUPAC_T165","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G90703NA\""},{"id":"GlycanIUPAC_T166","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G90703NA\""},{"id":"GlycanIUPAC_T167","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G90703NA\""},{"id":"GlycanIUPAC_T168","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G90703NA\""},{"id":"GlycanIUPAC_T169","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G01591HR\""},{"id":"GlycanIUPAC_T170","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G01591HR\""},{"id":"GlycanIUPAC_T171","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G01591HR\""},{"id":"GlycanIUPAC_T172","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G01591HR\""},{"id":"GlycanIUPAC_T173","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G56520XN\""},{"id":"GlycanIUPAC_T174","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G56520XN\""},{"id":"GlycanIUPAC_T175","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G56520XN\""},{"id":"GlycanIUPAC_T176","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G56520XN\""},{"id":"GlycanIUPAC_T177","span":{"begin":841,"end":847},"obj":"\"http://rdf.glycoinfo.org/glycan/G81830JX\""},{"id":"GlycanIUPAC_T178","span":{"begin":963,"end":969},"obj":"\"http://rdf.glycoinfo.org/glycan/G81830JX\""},{"id":"GlycanIUPAC_T179","span":{"begin":1131,"end":1137},"obj":"\"http://rdf.glycoinfo.org/glycan/G81830JX\""},{"id":"GlycanIUPAC_T180","span":{"begin":1146,"end":1152},"obj":"\"http://rdf.glycoinfo.org/glycan/G81830JX\""},{"id":"GlycanIUPAC_T181","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G19151BJ\""},{"id":"GlycanIUPAC_T182","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G63727IH\""},{"id":"GlycanIUPAC_T183","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G05795KA\""},{"id":"GlycanIUPAC_T184","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G90812SJ\""},{"id":"GlycanIUPAC_T185","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G07124DB\""},{"id":"GlycanIUPAC_T186","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G20567HS\""},{"id":"GlycanIUPAC_T187","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G76159WX\""},{"id":"GlycanIUPAC_T188","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G40623UW\""},{"id":"GlycanIUPAC_T189","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G06447MP\""},{"id":"GlycanIUPAC_T190","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G62764VZ\""},{"id":"GlycanIUPAC_T191","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G21434YY\""},{"id":"GlycanIUPAC_T192","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G34267PK\""},{"id":"GlycanIUPAC_T193","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G30235PC\""},{"id":"GlycanIUPAC_T194","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G34820UC\""},{"id":"GlycanIUPAC_T195","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G95531HX\""},{"id":"GlycanIUPAC_T196","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G74672YG\""},{"id":"GlycanIUPAC_T197","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G54440FW\""},{"id":"GlycanIUPAC_T198","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G69182II\""},{"id":"GlycanIUPAC_T199","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G68961XX\""},{"id":"GlycanIUPAC_T200","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G11710CQ\""},{"id":"GlycanIUPAC_T201","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G70765DS\""},{"id":"GlycanIUPAC_T202","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G51661YR\""},{"id":"GlycanIUPAC_T203","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G18477VG\""},{"id":"GlycanIUPAC_T204","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G12337MH\""},{"id":"GlycanIUPAC_T205","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G40783MY\""},{"id":"GlycanIUPAC_T206","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G72867LN\""},{"id":"GlycanIUPAC_T207","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G79061OT\""},{"id":"GlycanIUPAC_T208","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G45970UX\""},{"id":"GlycanIUPAC_T209","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G93661NW\""},{"id":"GlycanIUPAC_T210","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G04988XL\""},{"id":"GlycanIUPAC_T211","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G85800UH\""},{"id":"GlycanIUPAC_T212","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G96338GZ\""},{"id":"GlycanIUPAC_T213","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G96296PU\""},{"id":"GlycanIUPAC_T214","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G59124WZ\""},{"id":"GlycanIUPAC_T215","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G57275GR\""},{"id":"GlycanIUPAC_T216","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G46105MZ\""},{"id":"GlycanIUPAC_T217","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G76001GO\""},{"id":"GlycanIUPAC_T218","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G89599ML\""},{"id":"GlycanIUPAC_T219","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G85872BD\""},{"id":"GlycanIUPAC_T220","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G69460YK\""},{"id":"GlycanIUPAC_T221","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G25240JV\""},{"id":"GlycanIUPAC_T222","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G80489OS\""},{"id":"GlycanIUPAC_T223","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G35726XK\""},{"id":"GlycanIUPAC_T224","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G11002JA\""},{"id":"GlycanIUPAC_T225","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G85388FO\""},{"id":"GlycanIUPAC_T226","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G20221JL\""},{"id":"GlycanIUPAC_T227","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G39026SP\""},{"id":"GlycanIUPAC_T228","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G66979UZ\""},{"id":"GlycanIUPAC_T229","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G68566EA\""},{"id":"GlycanIUPAC_T230","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G20255ZD\""},{"id":"GlycanIUPAC_T231","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G68860MF\""},{"id":"GlycanIUPAC_T232","span":{"begin":1140,"end":1143},"obj":"\"http://rdf.glycoinfo.org/glycan/G50413KD\""}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"NCBITAXON","denotations":[{"id":"T1","span":{"begin":105,"end":120},"obj":"OrganismTaxon"},{"id":"T2","span":{"begin":606,"end":611},"obj":"OrganismTaxon"},{"id":"T3","span":{"begin":738,"end":753},"obj":"OrganismTaxon"},{"id":"T4","span":{"begin":973,"end":978},"obj":"OrganismTaxon"}],"attributes":[{"id":"A1","pred":"db_id","subj":"T1","obj":"4922"},{"id":"A2","pred":"db_id","subj":"T2","obj":"9606"},{"id":"A3","pred":"db_id","subj":"T3","obj":"4922"},{"id":"A4","pred":"db_id","subj":"T4","obj":"9606"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}

{"project":"Anatomy-UBERON","denotations":[{"id":"T1","span":{"begin":1230,"end":1235},"obj":"Body_part"}],"attributes":[{"id":"A1","pred":"uberon_id","subj":"T1","obj":"http://purl.obolibrary.org/obo/GO_0005794"}],"text":"Engineering of an artificial glycosylation pathway blocked in core oligosaccharide assembly in the yeast Pichia pastoris: production of complex humanized glycoproteins with terminal galactose.\nA significant percentage of eukaryotic proteins contain posttranslational modifications, including glycosylation, which are required for biological function. However, the understanding of the structure-function relationships of N-glycans has lagged significantly due to the microheterogeneity of glycosylation in mammalian produced proteins. Recently we reported on the cellular engineering of yeast to replicate human N-glycosylation for the production of glycoproteins. Here we report the engineering of an artificial glycosylation pathway in Pichia pastoris blocked in dolichol oligosaccharide assembly. The PpALG3 gene encoding Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol mannosyltransferase was deleted in a strain that was previously engineered to produce hybrid GlcNAcMan(5)GlcNAc(2) human N-glycans. Employing this approach, combined with the use of combinatorial genetic libraries, we engineered P. pastoris strains that synthesize complex GlcNAc(2)Man(3)GlcNAc(2) N-glycans with striking homogeneity. Furthermore, through expression of a Golgi-localized fusion protein comprising UDP-glucose 4-epimerase and beta-1,4-galactosyl transferase activities we demonstrate that this structure is a substrate for highly efficient in vivo galactose addition. Taken together, these data demonstrate that the artificial in vivo glycoengineering of yeast represents a major advance in the production of glycoproteins and will emerge as a practical tool to systematically elucidate the structure-function relationship of N-glycans."}