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. 1978 Oct 15;176(1):283–294. doi: 10.1042/bj1760283

Studies on the assembly of the rat lens capsule. Biosynthesis and partial characterization of the collagenous components.

G Heathcote, C H Sear, M E Grant
PMCID: PMC1186227  PMID: 215131

Abstract

1. Isolated rat lens capsules synthesized hydroxy[3H]proline-containing polypeptides when incubated with [3H]proline. 2. The collagenous polypeptides synthesized during a 2 h incubation were analyzed by both gel-filtration chromatography and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and shown to have an apparent mol.wt. of approx. 180,000. 3. No evidence was obtained for conversion of these polypeptides into a lower-molecular-weight species in experiments where capsules were labelled for 2 h and chased with non-radioactive proline for up to 22 h. However, a time-dependent incorporation of the 180,000-mol.wt. species into a larger collagenous component was observed and this could be prevented by the inclusion of beta-aminopropionitrile in the incubation medium. 4. The radioactive components synthesized by the capsules correspond to subunits of the intact lens capsule and the direct incorporation of the polypeptide of mol.wt. 180,000 into deoxycholate-insoluble basement membrane was demonstrated.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ashton N. Vascular basement membrane changes in diabetic retinopathy. Montgomery lecture, 1973. Br J Ophthalmol. 1974 Apr;58(4):344–366. doi: 10.1136/bjo.58.4.344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bailey A. J., Robins S. P., Balian G. Biological significance of the intermolecular crosslinks of collagen. Nature. 1974 Sep 13;251(5471):105–109. doi: 10.1038/251105a0. [DOI] [PubMed] [Google Scholar]
  3. Burgeson R. E., El Adli F. A., Kaitila I. I., Hollister D. W. Fetal membrane collagens: identification of two new collagen alpha chains. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2579–2583. doi: 10.1073/pnas.73.8.2579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark C. C., Tomichek E. A., Koszalka T. R., Minor R. R., Kefalides N. A. The embryonic rat parietal yolk sac. The role of the parietal endoderm in the biosynthesis of basement membrane collagen and glycoprotein in vitro. J Biol Chem. 1975 Jul 10;250(13):5259–5267. [PubMed] [Google Scholar]
  5. Daniels J. R., Chu G. H. Basement membrane collagen of renal glomerulus. J Biol Chem. 1975 May 10;250(9):3531–3537. [PubMed] [Google Scholar]
  6. Dixit S. N. Isolation and characterization of two collagenous components from anterior lens capsule. FEBS Lett. 1978 Jan 1;85(1):153–157. doi: 10.1016/0014-5793(78)81269-1. [DOI] [PubMed] [Google Scholar]
  7. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  8. Freytag J. W., Ohno M., Hudson B. G. Bovine renal glomerular basement membrane. Assessment of proteolysis during isolation. Biochem Biophys Res Commun. 1976 Oct 4;72(3):796–802. doi: 10.1016/s0006-291x(76)80203-3. [DOI] [PubMed] [Google Scholar]
  9. Fukushi S., Spiro R. G. The lens capsule. Sugar and amino acid composition. J Biol Chem. 1969 Apr 25;244(8):2041–2048. [PubMed] [Google Scholar]
  10. Grant M. E., Harwood R., Williams I. F. The biosynthesis of basement-membrane collagen by isolated rat glomeruli. Eur J Biochem. 1975 Jun;54(2):531–540. doi: 10.1111/j.1432-1033.1975.tb04166.x. [DOI] [PubMed] [Google Scholar]
  11. Grant M. E., Jackson D. S. The biosynthesis of procollagen. Essays Biochem. 1976;12:77–113. [PubMed] [Google Scholar]
  12. Grant M. E., Kefalides N. A., Prockop D. J. The biosynthesis of basement membrane collagen in embryonic chick lens. I. Delay between the synthesis of polypeptide chains and the secretion of collagen by matrix-free cells. J Biol Chem. 1972 Jun 10;247(11):3539–3544. [PubMed] [Google Scholar]
  13. Grant M. E., Kefalides N. A., Prockop D. J. The biosynthesis of basement membrane collagen in embryonic chick lens. II. Synthesis of a precursor form by matrix-free cells and a time-dependent conversion to chains in intact lens. J Biol Chem. 1972 Jun 10;247(11):3545–3551. [PubMed] [Google Scholar]
  14. Harwood R., Merry A. H., Woolley D. E., Grant M. E., Jackson D. S. The disulphide-bonded nature of procollagen and the role of the extension peptides in the assembly of the molecule. Biochem J. 1977 Feb 1;161(2):405–418. doi: 10.1042/bj1610405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Houston L. L. Amino acid analysis of stained bands from polyacrylamide gels. Anal Biochem. 1971 Nov;44(1):81–88. doi: 10.1016/0003-2697(71)90348-4. [DOI] [PubMed] [Google Scholar]
  16. Howard B. V., Macarak E. J., Gunson D., Kefalides N. A. Characterization of the collagen synthesized by endothelial cells in culture. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2361–2364. doi: 10.1073/pnas.73.7.2361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hudson B. G., Spiro R. G. Fractionation of glycoprotein components of the reduced alkylated renal glomerular basement membrane. J Biol Chem. 1972 Jul 10;247(13):4239–4247. [PubMed] [Google Scholar]
  18. Hudson B. G., Spiro R. G. Studies on the native and reduced alkylated renal glomerular basement membrane. Solubility, subunit size, and reaction with cyanogen bromide. J Biol Chem. 1972 Jul 10;247(13):4229–4238. [PubMed] [Google Scholar]
  19. Johnson L. D., Warfel J. Isolation and characterization of an epithelial basement membrane glycoprotein from murine kidney and further characterization of an epithelial basement membrane glycoprotein secreted by murine teratocarcinoma cells in vitro. Biochim Biophys Acta. 1976 Dec 2;455(2):538–549. doi: 10.1016/0005-2736(76)90323-0. [DOI] [PubMed] [Google Scholar]
  20. Juva K., Prockop D. J. Modified procedure for the assay of H-3-or C-14-labeled hydroxyproline. Anal Biochem. 1966 Apr;15(1):77–83. doi: 10.1016/0003-2697(66)90249-1. [DOI] [PubMed] [Google Scholar]
  21. KUNITZ M. Crystalline desoxyribonuclease; isolation and general properties; spectrophotometric method for the measurement of desoxyribonuclease activity. J Gen Physiol. 1950 Mar;33(4):349–362. doi: 10.1085/jgp.33.4.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kefalides N. A., Cameron J. D., Tomichek E. A., Yanoff M. Biosynthesis of basement membrane collagen by rabbit corneal endothelium in vitro. J Biol Chem. 1976 Feb 10;251(3):730–733. [PubMed] [Google Scholar]
  23. Kefalides N. A. Isolation of a collagen from basement membranes containing three identical - chains. Biochem Biophys Res Commun. 1971 Oct 1;45(1):226–234. doi: 10.1016/0006-291x(71)90073-8. [DOI] [PubMed] [Google Scholar]
  24. Kefalides N. A. Structure and biosynthesis of basement membranes. Int Rev Connect Tissue Res. 1973;6:63–104. doi: 10.1016/b978-0-12-363706-2.50008-8. [DOI] [PubMed] [Google Scholar]
  25. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  26. Lembach K. J., Branson R. E., Hewgley P. B., Cunningham L. W. The synthesis of macromolecular 3-hydroxyproline by attaching and confluent cultures of human fibroblasts. Eur J Biochem. 1977 Jan;72(2):379–383. doi: 10.1111/j.1432-1033.1977.tb11262.x. [DOI] [PubMed] [Google Scholar]
  27. Meezan E., Hjelle J. T., Brendel K., Carlson E. C. A simple, versatile, nondisruptive method for the isolation of morphologically and chemically pure basement membranes from several tissues. Life Sci. 1975 Dec 1;17(11):1721–1732. doi: 10.1016/0024-3205(75)90119-8. [DOI] [PubMed] [Google Scholar]
  28. Miller E. J. Biochemical characteristics and biological significance of the genetically-distinct collagens. Mol Cell Biochem. 1976 Dec 10;13(3):165–192. doi: 10.1007/BF01731779. [DOI] [PubMed] [Google Scholar]
  29. Miller E. J., Finch J. E., Jr, Chung E., Butler W. T., Robertson P. B. Specific cleavage of the native type III collagen molecule with trypsin. Similarity of the cleavage products to collagenase-produced fragments and primary structure at the cleavage site. Arch Biochem Biophys. 1976 Apr;173(2):631–637. doi: 10.1016/0003-9861(76)90300-3. [DOI] [PubMed] [Google Scholar]
  30. Minor R. R., Clark C. C., Strause E. L., Koszalka T. R., Brent R. L., Kefalides N. A. Basement membrane procollagen is not converted to collagen in organ cultures of parietal yolk sac endoderm. J Biol Chem. 1976 Mar 25;251(6):1789–1794. [PubMed] [Google Scholar]
  31. Minor R. R., Hoch P. S., Koszalka T. R., Brent R. L., Kefalides N. A. Organ cultures of the embryonic rat parietal yolk sac. I. Morphologic and autoradiographic studies of the deposition of the collagen and noncollagen glycoprotein components of basement membrane. Dev Biol. 1976 Feb;48(2):344–364. doi: 10.1016/0012-1606(76)90096-8. [DOI] [PubMed] [Google Scholar]
  32. Oho M., Riquetti P., Hudson B. G. Bovine renal glomerular basement membrane. Isolation and characterization of a glycoprotein component. J Biol Chem. 1975 Oct 10;250(19):7780–7787. [PubMed] [Google Scholar]
  33. Olsen B. R., Alper R., Kefalides N. A. Structural characterization of a soluble fraction from lens-capsule basement membrane. Eur J Biochem. 1973 Oct 5;38(2):220–228. doi: 10.1111/j.1432-1033.1973.tb03053.x. [DOI] [PubMed] [Google Scholar]
  34. Orkin R. W., Gehron P., McGoodwin E. B., Martin G. R., Valentine T., Swarm R. A murine tumor producing a matrix of basement membrane. J Exp Med. 1977 Jan 1;145(1):204–220. doi: 10.1084/jem.145.1.204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. PIRIE A. Composition of ox lens capsule. Biochem J. 1951 Mar;48(3):368–371. doi: 10.1042/bj0480368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Price R. G., Spiro R. G. Studies on the metabolism of the renal glomerular basement membrane. Turnover measurements in the rat with the use of radiolabeled amino acids. J Biol Chem. 1977 Dec 10;252(23):8597–8602. [PubMed] [Google Scholar]
  37. Prockop D. J. Role of iron in the synthesis of collagen in connective tissue. Fed Proc. 1971 May-Jun;30(3):984–990. [PubMed] [Google Scholar]
  38. Sato T., Spiro R. G. Studies on the subunit composition of the renal glomerular basement membrane. J Biol Chem. 1976 Jul 10;251(13):4062–4070. [PubMed] [Google Scholar]
  39. Schwartz D., Veis A. Characterization of basement membrane collagen of bovine anterior lens capsule via segment-long-spacing crystallites and the specific cleavage of the collagen by pepsin. FEBS Lett. 1978 Jan 15;85(2):326–332. doi: 10.1016/0014-5793(78)80484-0. [DOI] [PubMed] [Google Scholar]
  40. Spiro R. G. Search for a biochemical basis of diabetic microangiopathy. Diabetologia. 1976 Mar;12(1):1–14. doi: 10.1007/BF01221959. [DOI] [PubMed] [Google Scholar]
  41. Spiro R. G. Studies on the renal glomerular basement membrane. Preparation and chemical composition. J Biol Chem. 1967 Apr 25;242(8):1915–1922. [PubMed] [Google Scholar]
  42. Tanzer M. L., Kefalides N. A. Collagen crosslinks: occurrence in basement membrane collagens. Biochem Biophys Res Commun. 1973 Apr 2;51(3):775–780. doi: 10.1016/0006-291x(73)91382-x. [DOI] [PubMed] [Google Scholar]
  43. Taylor S. A., Price R. G. Characterization of subunits of rat glomerular basement membrane. Biochem Soc Trans. 1977;5(1):236–238. doi: 10.1042/bst0050236. [DOI] [PubMed] [Google Scholar]
  44. Timpl R., Martin G. R., Bruckner P., Wick G., Wiedemann H. Nature of the collagenous protein in a tumor basement membrane. Eur J Biochem. 1978 Mar;84(1):43–52. doi: 10.1111/j.1432-1033.1978.tb12139.x. [DOI] [PubMed] [Google Scholar]
  45. Trelstad R. L., Kang A. H. Collagen heterogeneity in the avian eye: lens, vitreous body, cornea and sclera. Exp Eye Res. 1974 Apr;18(4):395–406. doi: 10.1016/0014-4835(74)90117-1. [DOI] [PubMed] [Google Scholar]
  46. Vracko R. Basal lamina scaffold-anatomy and significance for maintenance of orderly tissue structure. Am J Pathol. 1974 Nov;77(2):314–346. [PMC free article] [PubMed] [Google Scholar]
  47. Young R. W., Ocumpaugh D. E. Autoradiographic studies on the growth and development of the lens capsule in the rat. Invest Ophthalmol. 1966 Dec;5(6):583–589. [PubMed] [Google Scholar]

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