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PMID: 8039 Published · ppublish English Journal Article

The route of secretion of procollagen. The influence of alphaalpha'-bipyridyl, colchicine and antimycin A on the secretory process in embryonic-chick tendon and cartilage cells.

The Biochemical journal ·Vol. 156 ·No. 1 ·1976-04-15 ·Pages 81-90

Harwood R, Grant ME, Jackson DS

Abstract

I. Embryonic-chick tendon cells were pulse-labelled for 4 min with [14C]proline and the 14C-labelled polypeptides were chased with unlabelled proline for up to 30 min. Isolation of subcellular fractions during the chase period and their subsequent analysis for bacterial collagenase-susceptible 14C-labelled peptides demonstrated the transfer of procollagen polypeptides from rough to smooth microsomal fractions and thence to the extracellular medium. Parallel analyses of Golgi-enriched fractions indicated the involvement of this organelle in the secretory pathway of procollagen. Sodium dodecylsulphate/polyacrylamide-gel electrophoresis of the 14C-labelled polypeptides present in the Golgi-enriched fractions demonstrated that the procollagen polypeptides were all present as disulphide-linked pro-gamma components. 2. When similar kinetic studies of the intracellular transport of procollagen were conducted with embryonic-chick cartilage cells almost identical results were obtained, but the rate of translocation of cartilage procollagen was significantly slower than that observed for tendon procollagen. 3. When hydroxylation of procollagen polypeptides was inhibited by alphaalpha'-bipyridyl, the nascent polypeptides accumulated in the rough microsomal fraction. 4. When cells were pulse-labelled for 4min with [14C)proline and the label was chased in the presence of colchicine, secretion of procollagen was inhibited and an intracellular accumulation of procollagen 14C-labelled polypeptides was observed in the Golgi-enriched fractions. 5. The energy-dependence of the intracellular transport of procollagen was demonstrated in experiments in which antimycin A was found to inhibit the transfer of procollagen polypeptides from rough to smooth endoplasmic reticulum. 6. It is concluded that procollagen follows the classical route of secretion taken by other extracellular proteins.

MeSH Terms
2,2'-Dipyridyl/pharmacology Animals Antimycin A/pharmacology Cartilage/metabolism Chick Embryo Colchicine/pharmacology Collagen/biosynthesis,metabolism Hydroxylation In Vitro Techniques Proline/metabolism Protein Precursors/metabolism Pyridines/pharmacology Subcellular Fractions/metabolism Tendons/metabolism
Chemicals
Protein Precursors Pyridines 2,2'-Dipyridyl Antimycin A Collagen Proline Colchicine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Harwood R
Grant M E
Jackson D S
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55 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1560081
Published
1976-04-15
Pages
81-90
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1163719
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1560081 PMC Full Text View journal details
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