PubMed:2505925 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PubMed/sourceid/2505925","sourcedb":"PubMed","sourceid":"2505925","source_url":"http://www.ncbi.nlm.nih.gov/pubmed/2505925","text":"Purification of endo-N-acetyl-beta-D-glucosaminidase H by substrate-affinity chromatography.\nEndo-N-acetyl-beta-D-glucosaminidase H (Endo H) was purified to homogeneity (3000-fold) from a culture filtrate to Streptomyces plicatus. The key step was substrate-affinity chromatography, which afforded a 1000-fold purification and yielded a protease- and exoglycosidase-free preparation of Endo H. Proteins from the crude sample were applied to the substrate-affinity column, consisting of yeast-invertase glycopeptides bound to Sepharose-immobilized concanavalin A. After washing off the unbound proteins, Endo H was quantitatively eluted by methyl alpha-D-mannopyranoside. Various conditions were tested to achieve an optimal binding of Endo H to this substrate-affinity gel. After substrate-affinity chromatography, Endo H was separated from the coeluted gylcopeptide substrate and some protein impurities by gel filtration and hydrophobic chromatography.","tracks":[{"project":"sentences","denotations":[{"id":"TextSentencer_T1","span":{"begin":0,"end":92},"obj":"Sentence"},{"id":"TextSentencer_T2","span":{"begin":93,"end":230},"obj":"Sentence"},{"id":"TextSentencer_T3","span":{"begin":231,"end":393},"obj":"Sentence"},{"id":"TextSentencer_T4","span":{"begin":394,"end":562},"obj":"Sentence"},{"id":"TextSentencer_T5","span":{"begin":563,"end":670},"obj":"Sentence"},{"id":"TextSentencer_T6","span":{"begin":671,"end":773},"obj":"Sentence"},{"id":"TextSentencer_T7","span":{"begin":774,"end":954},"obj":"Sentence"},{"id":"T1","span":{"begin":0,"end":92},"obj":"Sentence"},{"id":"T2","span":{"begin":93,"end":230},"obj":"Sentence"},{"id":"T3","span":{"begin":231,"end":393},"obj":"Sentence"},{"id":"T4","span":{"begin":394,"end":562},"obj":"Sentence"},{"id":"T5","span":{"begin":563,"end":670},"obj":"Sentence"},{"id":"T6","span":{"begin":671,"end":773},"obj":"Sentence"},{"id":"T7","span":{"begin":774,"end":954},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"attributes":[{"subj":"TextSentencer_T1","pred":"source","obj":"sentences"},{"subj":"TextSentencer_T2","pred":"source","obj":"sentences"},{"subj":"TextSentencer_T3","pred":"source","obj":"sentences"},{"subj":"TextSentencer_T4","pred":"source","obj":"sentences"},{"subj":"TextSentencer_T5","pred":"source","obj":"sentences"},{"subj":"TextSentencer_T6","pred":"source","obj":"sentences"},{"subj":"TextSentencer_T7","pred":"source","obj":"sentences"},{"subj":"T1","pred":"source","obj":"sentences"},{"subj":"T2","pred":"source","obj":"sentences"},{"subj":"T3","pred":"source","obj":"sentences"},{"subj":"T4","pred":"source","obj":"sentences"},{"subj":"T5","pred":"source","obj":"sentences"},{"subj":"T6","pred":"source","obj":"sentences"},{"subj":"T7","pred":"source","obj":"sentences"}]},{"project":"NCBITAXON","denotations":[{"id":"T1","span":{"begin":208,"end":229},"obj":"OrganismTaxon"}],"attributes":[{"id":"A1","pred":"db_id","subj":"T1","obj":"1922"},{"subj":"T1","pred":"source","obj":"NCBITAXON"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"sentences","color":"#ec93c5","default":true},{"id":"NCBITAXON","color":"#93dfec"}]}]}}