Incorporation of sulphate into type III procollagen by cultured human fibroblasts. Identification of tyrosine O-sulphate
Open Access
- 1 January 1986
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 154 (1), 219-224
- https://doi.org/10.1111/j.1432-1033.1986.tb09382.x
Abstract
Confluent cultures of normal human skin fibroblasts were labelled overnight wtih [35S]sulphate, and the incorporation of the isotope into type III procollagen, secreted into the medium, was verified by radioimmunoassay and immunoprecipitation after removing the heavily sulphated proteoglycans by anion-exchange chromatography. Type III procollagen and its pro and pN .alpha. chains were visualized in fluorographs of the immunoprecipitates. The labelled procollagen could be isolated by a combination of ion-exchange chromatography and gel filtration and was found to contain tyrosine O-sulphate, which was identified by thin-layer electrophoresis after Ba(OH)2 hydrolysis. The regions sulphated in the type III procollagen molecule were susceptible to pepsin digestion. Digestion with purified bacterial collagenase at +37.degree. C produced a labelled fragment that was recognized by antibodies against the aminoterminal propeptide of type III procollagen, indicating that the sulphated tyrosine residues are located either in this propeptide or in the non-helical telopeptide region of the type III collagen molecule proper. Sulphation of tyrosine residues is a new post-translational modification in procollagen, which could be involved in the regulation of the processing of type III procollagen into collagen and thus affect the formation of collagen fibres.This publication has 28 references indexed in Scilit:
- The complete amino acid sequence of hirudin, a thrombin specific inhibitorFEBS Letters, 1984
- Synthesis and localization of two sulphated glycoproteins associated with basement membranes and the extracellular matrix.The Journal of cell biology, 1982
- Heterogeneity of the antigens related to the aminoterminal propeptide of type III procollagen in human serumClinica Chimica Acta; International Journal of Clinical Chemistry, 1982
- Studies on the Biosynthesis of Laminin by Murine Parietal Endoderm CellsEuropean Journal of Biochemistry, 1981
- The Biosynthesis of Collagen and Its DisordersNew England Journal of Medicine, 1979
- Isolation of Crosslinked Peptides from Insoluble Human LeiomyomaEuropean Journal of Biochemistry, 1979
- Biosynthesis of ProcollagenAnnual Review of Biochemistry, 1978
- The Diagnostic Application of Specific Antiprocollagen SeraInternational Archives of Allergy and Immunology, 1978
- Characterization of the Amino-Terminal Segment in Type III ProcollagenEuropean Journal of Biochemistry, 1976
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970