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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
References (55)
55 references, click to expand
  1. Interaction of drugs with microtubule proteins.
    Fed Proc. 1974 Feb;33(2):158-66 PMID: 4855812
  2. Ferritin-conjugated antibodies used for labeling of organelles involved in the cellular synthesis and transport of procollagen.
    Proc Natl Acad Sci U S A. 1974 May;71(5):2033-7 PMID: 4525314
  3. Collagen formation in developing molar teeth of rats.
    J Ultrastruct Res. 1967 Dec;21(5):383-414 PMID: 5590726
  4. Intracellular accumulation of protocollagen and extrusion of collagen by embryonic cartilage cells.
    J Cell Biol. 1968 Sep;38(3):523-37 PMID: 5664222
  5. The biosynthesis of collagen. 1.
    N Engl J Med. 1972 Jan 27;286(4):194-9 PMID: 4257078
  6. Hydroxyproline stabilizes the triple helix of chick tendon collagen.
    Biochem Biophys Res Commun. 1973 May 1;52(1):106-14 PMID: 4576266
  7. Functions of polyribosomes attached to membranes of animal cells.
    FEBS Lett. 1970 Mar 16;7(1):1-7 PMID: 11947416
  8. Presumptive mRNA for procollagen: occurrence in membrane bound ribosomes of embryonic chick tendon fibroblasts.
    FEBS Lett. 1974 Apr 15;41(1):85-8 PMID: 4859449
  9. The sub-cellular location of inter-chain disulfide bond formation during procollagen biosynthesis by embryonic chick tendon cells.
    Biochem Biophys Res Commun. 1973 Dec 19;55(4):1188-96 PMID: 4771992
  10. Evidence for the participation of the Golgi apparatus in the intracellular transport of nascent albumin in the liver cell.
    J Cell Biol. 1970 Dec;47(3):555-67 PMID: 5497537
  11. Intracellular localization of liver sugar nucleotide glycoprotein glycosyltransferases in a Golgi-rich fraction.
    J Biol Chem. 1970 Mar 10;245(5):1090-100 PMID: 4392041
  12. Effect of urea on trypsin and alpha-chymotrypsin.
    Nature. 1956 Mar 10;177(4506):471-3 PMID: 13309338
  13. Controlled proteolysis of nascent polypeptides in rat liver cell fractions. II. Location of the polypeptides in rough microsomes.
    J Cell Biol. 1970 Apr;45(1):146-57 PMID: 5458993
  14. The biosynthesis of collagen. 3.
    N Engl J Med. 1972 Feb 10;286(6):291-300 PMID: 4550295
  15. Colchicine-binding protein and the secretion of thyroid hormone.
    J Cell Biol. 1972 Jul;54(1):157-65 PMID: 4338961
  16. Intracellular transport of secretory proteins in the pancreatic exocrine cell. IV. Metabolic requirements.
    J Cell Biol. 1968 Dec;39(3):589-603 PMID: 5699933
  17. Evidence for a role of the microtubular system in the secretion of newly synthesized albumin and other proteins by the liver.
    J Clin Invest. 1974 Jun;53(6):1512-7 PMID: 4830218
  18. Wound healing and collagen formation. V. Quantitative electron microscope radioautographic observations of proline-H3 utilization by fibroblasts.
    J Cell Biol. 1965 Oct;27(1):83-106 PMID: 5859922
  19. Secretion of procollagen: evidence for the transfer of nascent polypeptides across microsomal membranes of tendon cells.
    Biochem Biophys Res Commun. 1974 Aug 5;59(3):947-54 PMID: 4472297
  20. Intracellular transport of secretory proteins in the pancreatic exocrine cell. I. Role of the peripheral elements of the Golgi complex.
    J Cell Biol. 1967 Aug;34(2):577-96 PMID: 6035647
  21. Inhibition of collagen secretion from bone and cultured fibroblasts by microtubular disruptive drugs.
    Proc Natl Acad Sci U S A. 1972 Apr;69(4):892-6 PMID: 4502941
  22. Accumulation of collagen-containing vacuoles in osteoblasts after administration of colchicine.
    Cell Tissue Res. 1975;157(3):353-65 PMID: 164288
  23. AN AUTORADIOGRAPHIC AND ELECTRON MICROSCOPIC STUDY OF COLLAGEN SYNTHESIS IN DIFFERENTIATING CARTILAGE.
    Z Zellforsch Mikrosk Anat. 1963 Oct 8;61:110-44 PMID: 14095095
  24. Controlled proteolysis of nascent polypeptides in rat liver cell fractions. I. Location of the polypeptides within ribosomes.
    J Cell Biol. 1970 Apr;45(1):130-45 PMID: 5458992
  25. Influence of ascorbic acid on ribosomal patterns and collagen biosynthesis in healing wounds of scorbutic guinea pigs.
    Biochem J. 1974 Sep;142(3):641-51 PMID: 4464846
  26. Studies on acute phase proteins of rat serum. IV. Pathway of secretion of albumin and alpha1-acid glycoprotein from liver.
    Can J Biochem. 1973 Sep;51(9):1281-91 PMID: 4742881
  27. The metabolic requirements for transcellular movement and secretion of collagen.
    J Biol Chem. 1975 Jul 10;250(13):4841-7 PMID: 1150643
  28. Synthesis and secretion of under-hydroxylated procollagen at various temperatures by cells subject to temporary anoxia.
    Biochem Biophys Res Commun. 1974 Sep 9;60(1):414-23 PMID: 4472803
  29. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis.
    J Biol Chem. 1969 Aug 25;244(16):4406-12 PMID: 5806584
  30. The biosynthesis of collagen. 2.
    N Engl J Med. 1972 Feb 3;286(5):242-9 PMID: 4332072
  31. The synthesis and secretion of cartilage procollagen.
    Biochem J. 1975 Apr;148(1):129-38 PMID: 1156392
  32. Cell-free collagen synthesis on membrane-bound polysomes of chick embryo connective tissue and the localization of prolyl hydroxylase on the polysome-membrane complex.
    J Biol Chem. 1973 Sep 25;248(18):6514-21 PMID: 4354214
  33. The thermal transition of a non-hydroxylated form of collagen. Evidence for a role for hydroxyproline in stabilizing the triple-helix of collagen.
    Biochem Biophys Res Commun. 1973 May 1;52(1):115-20 PMID: 4712181
  34. Time lag in the secretion of collagen by matrix-free tendon cells and inhibition of the secretory process by colchicine and vinblastine.
    Biochim Biophys Acta. 1972 Apr 21;264(2):375-82 PMID: 4337621
  35. Characterization of the colchicine binding of membrane fractions from rat and mouse liver.
    J Cell Biol. 1974 Jan;60(1):297-303 PMID: 4809244
  36. Studies on the glycosylation of hydroxylysine residues during collagen biosynthesis and the subcellular localization of collagen galactosyltransferase and collagen glucosyltransferase in tendon and cartilage cells.
    Biochem J. 1975 Nov;152(2):291-302 PMID: 1220686
  37. Intracellular hydroxylation of non-helical protocollagen to form triple-helical procollagen and subsequent secretion of the molecule.
    Eur J Biochem. 1974 Apr 1;43(2):221-30 PMID: 4365178
  38. Studies on cartilage. III. The occurrence of collagen within vacuoles of the golgi apparatus.
    J Cell Biol. 1962 Mar;12:599-613 PMID: 13911612
  39. Colchicine inhibition of plasma protein release from rat hepatocytes.
    J Cell Biol. 1975 Jul;66(1):42-59 PMID: 1141379
  40. Effects of antimicrotubular agents on the secretion of collagen. A biochemical and morphological study.
    J Cell Biol. 1974 Aug;62(2):390-405 PMID: 4426914
  41. Location of procollagen in chick corneal and tendon fibroblasts with ferritin-conjugated antibodies.
    J Cell Biol. 1975 Apr;65(1):75-87 PMID: 1168646
  42. Collagen polypeptides: normal release from polysomes in the absence of proline hydroxylation.
    Science. 1971 Aug 20;173(3998):723-5 PMID: 5568504
  43. [Quantitative electron microscopic autoradiographic study of osteogenesis with tritiated proline].
    Z Zellforsch Mikrosk Anat. 1969;99(1):121-33 PMID: 5810774
  44. Biosynthesis of cartilage procollagen.
    Eur J Biochem. 1973 May;35(1):159-66 PMID: 4351528
  45. H3-proline incorporation into cartilage: electron microscope autoradiographic observations.
    J Morphol. 1968 Apr;124(4):387-421 PMID: 5666527
  46. Collagen formation--observations on its intracellular packaging and transport.
    Z Zellforsch Mikrosk Anat. 1972;129(4):455-70 PMID: 4339506
  47. Direct evidence for a colchicine-induced impairment in the mobility of membrane components.
    Science. 1973 Dec 14;182(4117):1136-8 PMID: 4201508
  48. Secretion of collagen by corneal epithelium. I. Morphology of the collagenous products produced by isolated epithelia grown on frozen-killed lens.
    J Cell Biol. 1973 Apr;57(1):190-213 PMID: 4347977
  49. Intracellular aspects of the process of protein synthesis.
    Science. 1975 Aug 1;189(4200):347-58 PMID: 1096303
  50. The preparation of a bacterial collagenase containing negligible non-specific protease activity.
    Prep Biochem. 1975;5(3):229-45 PMID: 170602
  51. Synthesis, migration, and release of precursor collagen by odontoblasts as visualized by radioautography after (3H)proline administration.
    J Cell Biol. 1974 Jan;60(1):92-127 PMID: 4129079
  52. Microtubules in transcellular movement of procollagen.
    Nat New Biol. 1972 Aug 30;238(87):257-60 PMID: 4342528
  53. Colchicine-induced inhibition of very low density lipoprotein release by rat liver in vivo.
    Biochim Biophys Acta. 1973 Apr 13;306(1):142-7 PMID: 4350003
  54. Further characterization of embryonic tendon fibroblasts and the use of immunoferritin techniques to study collagen biosynthesis.
    J Cell Biol. 1975 Feb;64(2):340-55 PMID: 163830
  55. STUDIES ON THE STRUCTURAL AND ENZYMIC ORGANIZATION OF THE MEMBRANOUS ELEMENTS OF LIVER MICROSOMES.
    Acta Pathol Microbiol Scand Suppl. 1963;:SUPPL166:1-94 PMID: 14080472
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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