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Article: Comprehensive analysis of collagen metabolism in vitro using [43H]/[14C]proline dual-labeling and polyacrylamide gel electrophoresis

TitleComprehensive analysis of collagen metabolism in vitro using [43H]/[14C]proline dual-labeling and polyacrylamide gel electrophoresis
Authors
Keywordscell culture
collagen metabolism
electrophoresis
proline hydroxylation
Issue Date1988
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/yabio
Citation
Analytical Biochemistry, 1988, v. 168 n. 1, p. 171-175 How to Cite?
AbstractA method to simultaneously quantify the production, secretion, and prolyl hydroxylation of individual types of collagen in cell culture samples has been developed. Collagens were biosynthetically labeled with a mixture of [14C]proline and [4-3H]proline. The labeled collagens were isolated and their component α-chains were resolved by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Migration of the collagen α-chains was determined by fluorography, and radioactivity in excised bands was quantified by scintillation counting. [14C]proline labeling of collagen chains was used to determine the production and secretion of the different types of collagen. The ratios of the component α1(I) and α2(I) chains of type I collagen were also determined in this way. Prolyl hydroxylation of collagen α-chains was readily determined by measurement of their 3H:14C ratios. Following 4-hydroxylation, 3H was lost from the [4-3H]proline with alteration of this ratio. This dual-labeling method is suitable for the comprehensive analysis of collagen metabolism in multiple samples.
Persistent Identifierhttp://hdl.handle.net/10722/147329
ISSN
2023 Impact Factor: 2.6
2023 SCImago Journal Rankings: 0.493
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorBateman, JFen_US
dc.contributor.authorHarley, Ven_US
dc.contributor.authorChan, Den_US
dc.contributor.authorCole, WGen_US
dc.date.accessioned2012-05-29T06:02:57Z-
dc.date.available2012-05-29T06:02:57Z-
dc.date.issued1988en_US
dc.identifier.citationAnalytical Biochemistry, 1988, v. 168 n. 1, p. 171-175en_US
dc.identifier.issn0003-2697en_US
dc.identifier.urihttp://hdl.handle.net/10722/147329-
dc.description.abstractA method to simultaneously quantify the production, secretion, and prolyl hydroxylation of individual types of collagen in cell culture samples has been developed. Collagens were biosynthetically labeled with a mixture of [14C]proline and [4-3H]proline. The labeled collagens were isolated and their component α-chains were resolved by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Migration of the collagen α-chains was determined by fluorography, and radioactivity in excised bands was quantified by scintillation counting. [14C]proline labeling of collagen chains was used to determine the production and secretion of the different types of collagen. The ratios of the component α1(I) and α2(I) chains of type I collagen were also determined in this way. Prolyl hydroxylation of collagen α-chains was readily determined by measurement of their 3H:14C ratios. Following 4-hydroxylation, 3H was lost from the [4-3H]proline with alteration of this ratio. This dual-labeling method is suitable for the comprehensive analysis of collagen metabolism in multiple samples.en_US
dc.languageengen_US
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/yabioen_US
dc.relation.ispartofAnalytical Biochemistryen_US
dc.subjectcell culture-
dc.subjectcollagen metabolism-
dc.subjectelectrophoresis-
dc.subjectproline hydroxylation-
dc.subject.meshCarbon Radioisotopes - Diagnostic Useen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshCollagen - Analysis - Biosynthesis - Secretionen_US
dc.subject.meshElectrophoresis, Polyacrylamide Gelen_US
dc.subject.meshFibroblasts - Metabolismen_US
dc.subject.meshHumansen_US
dc.subject.meshHydroxylationen_US
dc.subject.meshIsotope Labelingen_US
dc.subject.meshMethylationen_US
dc.subject.meshProline - Diagnostic Use - Metabolismen_US
dc.subject.meshTritium - Diagnostic Useen_US
dc.titleComprehensive analysis of collagen metabolism in vitro using [43H]/[14C]proline dual-labeling and polyacrylamide gel electrophoresisen_US
dc.typeArticleen_US
dc.identifier.emailChan, D:chand@hkucc.hku.hken_US
dc.identifier.authorityChan, D=rp00540en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/0003-2697(88)90025-5-
dc.identifier.pmid3364711en_US
dc.identifier.scopuseid_2-s2.0-0023848205en_US
dc.identifier.volume168en_US
dc.identifier.issue1en_US
dc.identifier.spage171en_US
dc.identifier.epage175en_US
dc.identifier.isiWOS:A1988L731600024-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridBateman, JF=16135557700en_US
dc.identifier.scopusauthoridHarley, V=35465928300en_US
dc.identifier.scopusauthoridChan, D=7402216545en_US
dc.identifier.scopusauthoridCole, WG=7201518727en_US
dc.identifier.issnl0003-2697-

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