Synthesis of procollagen by matrix-free cells from embryonic-chick arteries
- 15 September 1977
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 166 (3), 437-446
- https://doi.org/10.1042/bj1660437
Abstract
Cells were isolated from the major arteries of 17-day chick embryos by digestion of the tissue with collagenase and trypsin. The cells, when examined immediately after isolation, exhibited a high degree of viability and they were shown to synthesize and secrete procollagen at a high and constant rate for several hours when incubated in suspension in modified Krebs medium. Continuous labelling of the cells with [14C]proline demonstrated a lag of about 30min between the time at which the synthesis of non-diffusible peptide-bound hydroxy[14C]proline became linear and the time at which its secretion into the medium became linear. This lag time compares with that of 18min observed for freshly isolated matrix-free cells from embryonic-chick tendon, which synthesize and secrete the same type of collagen. Gel-filtration chromatography and polyacrylamide-gel electrophoresis indicated that the collagenous polypeptides secreted into the medium were in the precursor form, known as procollagen, and that the constituent pro-α-chains were linked by interchain disulphide bonds and were also in a triple-helical conformation. Characterization of the secreted procollagen by gel-filtration chromatography, polyacrylamide-gel electrophoresis, DEAE-agarose chromatography, and polyacrylamide-gel electrophoresis of peptides obtained by CNBr cleavage, indicated that the predominant form was type-I procollagen. This work extends the range of freshly isolated matrix-free cell systems, which have been characterized for use in studies on the biosynthesis and secretion of procollagen, and it indicates differences in the rates of secretion of procollagen in different cell types secreting the same type of procollagen.This publication has 31 references indexed in Scilit:
- Identification of a major extracellular non-collagenous glycoprotein synthesised by human skin fibroblasts in cultureBiochemical and Biophysical Research Communications, 1976
- Biosynthesis and secretion of tropoelastin by chick aorta cellsBiochemical and Biophysical Research Communications, 1976
- The separation of two soft-tissue collagens by covalent chromatographyFEBS Letters, 1976
- ProcollagenPublished by Wiley ,1975
- Human aorta collagens: Evidence for three distinct speciesBiochemical and Biophysical Research Communications, 1974
- Collagen Polymorphism: Characterization of Molecules with the Chain Composition [α1(III)] 3 in Human TissuesScience, 1974
- Time lag in the secretion of collagen by matrix-free tendon cells and inhibition of the secretory process by colchicine and vinblastineBiochimica et Biophysica Acta (BBA) - General Subjects, 1972
- Identification of a soluble intermediate during synthesis of elastin by embryonic chick aortaeFEBS Letters, 1972
- Further evidence for a transport form of collagen. Its extrusion and extracellular conversion to tropocollagen in embryonic tendonFEBS Letters, 1971
- Purification and partial characterization of a soluble elastin-like protein from copper-deficient procine aortaBiochemistry, 1969