PMC:1914094 / 11430-12623 JSONTXT

{"project":"2_test","denotations":[{"id":"17557078-12089508-27549121","span":{"begin":376,"end":380},"obj":"12089508"}],"text":"Kinetic trapping can be applied to detect Trx1 interactions on the cell surface\nHaving established the Trx1 kinetic trapping approach for soluble target proteins, we asked whether the kinetic trapping technique can also be applied to the surface of intact cells in culture. Given previous indications of disulfide bond exchange between CD4 and wild-type Trx1 (Matthias et al, 2002), we asked whether kinetic trapping would enable us to detect this interaction on the surface of the CD4 positive promyelocytic cell line U937. In brief, we allowed mutant Trx1 to interact with the surface of live cells, removed unreacted oxidoreductase by washing and collected disulfide-linked Trx1 complexes from cellular lysates by streptavidin (SAv) affinity purification. We found that cell surface CD4 forms a mixed disulfide with exogenously added Trx1(CSAAA) (Supplementary Figure S1, left panel), which could be dissociated by DTT treatment (Supplementary Figure S1, right panel). This result confirmed that kinetic trapping can indeed be used to identify specific Trx1-reactive cell surface proteins and should therefore allow de novo identification of previously unknown cell surface target proteins."}

{"project":"WikiPainGoldStandard","denotations":[{"id":"T59","span":{"begin":314,"end":318},"obj":"BINDING"},{"id":"T60","span":{"begin":561,"end":569},"obj":"BINDING"},{"id":"T61","span":{"begin":682,"end":691},"obj":"BINDING"},{"id":"T62","span":{"begin":804,"end":813},"obj":"BINDING"},{"id":"T63","span":{"begin":336,"end":339},"obj":"Protein entrezID:920"},{"id":"T64","span":{"begin":354,"end":358},"obj":"Protein entrezID:7295"},{"id":"T65","span":{"begin":553,"end":557},"obj":"Protein entrezID:7295"},{"id":"T66","span":{"begin":677,"end":681},"obj":"Protein entrezID:7295"},{"id":"T67","span":{"begin":786,"end":789},"obj":"Protein entrezID:920"},{"id":"T68","span":{"begin":837,"end":841},"obj":"Protein entrezID:7295"}],"relations":[{"id":"R44","pred":"REFERENCED THEME","subj":"T59","obj":"T64"},{"id":"R45","pred":"REPORTED THEME","subj":"T60","obj":"T65"},{"id":"R46","pred":"REPORTED THEME","subj":"T61","obj":"T66"},{"id":"R47","pred":"REPORTED THEME","subj":"T62","obj":"T68"},{"id":"R48","pred":"REFERENCED CAUSE","subj":"T63","obj":"T59"},{"id":"R49","pred":"REPORTED CAUSE","subj":"T67","obj":"T62"}],"text":"Kinetic trapping can be applied to detect Trx1 interactions on the cell surface\nHaving established the Trx1 kinetic trapping approach for soluble target proteins, we asked whether the kinetic trapping technique can also be applied to the surface of intact cells in culture. Given previous indications of disulfide bond exchange between CD4 and wild-type Trx1 (Matthias et al, 2002), we asked whether kinetic trapping would enable us to detect this interaction on the surface of the CD4 positive promyelocytic cell line U937. In brief, we allowed mutant Trx1 to interact with the surface of live cells, removed unreacted oxidoreductase by washing and collected disulfide-linked Trx1 complexes from cellular lysates by streptavidin (SAv) affinity purification. We found that cell surface CD4 forms a mixed disulfide with exogenously added Trx1(CSAAA) (Supplementary Figure S1, left panel), which could be dissociated by DTT treatment (Supplementary Figure S1, right panel). This result confirmed that kinetic trapping can indeed be used to identify specific Trx1-reactive cell surface proteins and should therefore allow de novo identification of previously unknown cell surface target proteins."}

{"project":"MyTest","denotations":[{"id":"17557078-12089508-27549121","span":{"begin":376,"end":380},"obj":"12089508"}],"namespaces":[{"prefix":"_base","uri":"https://www.uniprot.org/uniprot/testbase"},{"prefix":"UniProtKB","uri":"https://www.uniprot.org/uniprot/"},{"prefix":"uniprot","uri":"https://www.uniprot.org/uniprotkb/"}],"text":"Kinetic trapping can be applied to detect Trx1 interactions on the cell surface\nHaving established the Trx1 kinetic trapping approach for soluble target proteins, we asked whether the kinetic trapping technique can also be applied to the surface of intact cells in culture. Given previous indications of disulfide bond exchange between CD4 and wild-type Trx1 (Matthias et al, 2002), we asked whether kinetic trapping would enable us to detect this interaction on the surface of the CD4 positive promyelocytic cell line U937. In brief, we allowed mutant Trx1 to interact with the surface of live cells, removed unreacted oxidoreductase by washing and collected disulfide-linked Trx1 complexes from cellular lysates by streptavidin (SAv) affinity purification. We found that cell surface CD4 forms a mixed disulfide with exogenously added Trx1(CSAAA) (Supplementary Figure S1, left panel), which could be dissociated by DTT treatment (Supplementary Figure S1, right panel). This result confirmed that kinetic trapping can indeed be used to identify specific Trx1-reactive cell surface proteins and should therefore allow de novo identification of previously unknown cell surface target proteins."}