Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1981 Mar 1;193(3):749–755. doi: 10.1042/bj1930749

Isolation and characterization of pepsin fragments of laminin from human placental and renal basement membranes.

L Risteli, R Timpl
PMCID: PMC1162663  PMID: 6796051

Abstract

The presence of laminin in authentic basement membranes was examined at the level of a large pepsin-resistant fragment P1. This strongly antigenic fragment has been recently isolated from a mouse tumour basement membrane. By using antibodies to mouse laminin P1 for identification it was possible to isolate a homologous fragment P1 (Mr about 250 000) and a related component Pa (Mr about 70 000--90 000) from pepsin digests of human placenta and kidney. The fragments were in half-cystine (90--130 residues/1000) and carbohydrate and showed strong binding to concanavalin A. Reduction of disulphide bonds produced several smaller peptide chains, indicating a complex pepsin cleavage. Immunological assays demonstrated partial antigenic identity between laminin fragments obtained from mouse and human tissue, and suggested that fragment Pa may originate from a protein not completely identical with laminin. The results showed that laminin is an abundant component of tissue rich in basement membranes, which has been previously suggested by immunohistological studies.

Full text

PDF
750

Images in this article

750Fig. 1.
on p.750
752Fig. 4.
on p.752

Selected References

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

  1. Alitalo K., Kurkinen M., Vaheri A., Krieg T., Timpl R. Extracellular matrix components synthesized by human amniotic epithelial cells in culture. Cell. 1980 Apr;19(4):1053–1062. doi: 10.1016/0092-8674(80)90096-3. [DOI] [PubMed] [Google Scholar]
  2. Alitalo K., Vaheri A., Krieg T., Timpl R. Biosynthesis of two subunits of type IV procollagen and of other basement membrane proteins by a human tumor cell line. Eur J Biochem. 1980 Aug;109(1):247–255. doi: 10.1111/j.1432-1033.1980.tb04790.x. [DOI] [PubMed] [Google Scholar]
  3. Anttinen H., Myllylä R., Kivirikko K. I. Hydrophobic and carbohydrate-recognition chromatographies of collagen glucosyltransferase. Eur J Biochem. 1977 Aug 15;78(1):11–17. doi: 10.1111/j.1432-1033.1977.tb11708.x. [DOI] [PubMed] [Google Scholar]
  4. Bailey A. J., Sims T. J., Duance V. C., Light N. D. Partial characterization of a second basement membrane collagen in human placenta. Evidence for the existence of two type IV collagen molecules. FEBS Lett. 1979 Mar 15;99(2):361–366. doi: 10.1016/0014-5793(79)80992-8. [DOI] [PubMed] [Google Scholar]
  5. Chung A. E., Jaffe R., Freeman I. L., Vergnes J. P., Braginski J. E., Carlin B. Properties of a basement membrane-related glycoprotein synthesized in culture by a mouse embryonal carcinoma-derived cell line. Cell. 1979 Feb;16(2):277–287. doi: 10.1016/0092-8674(79)90005-9. [DOI] [PubMed] [Google Scholar]
  6. Ekblom P., Alitalo K., Vaheri A., Timpl R., Saxén L. Induction of a basement membrane glycoprotein in embryonic kidney: possible role of laminin in morphogenesis. Proc Natl Acad Sci U S A. 1980 Jan;77(1):485–489. doi: 10.1073/pnas.77.1.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. FRANCOIS C., MARSHALL R. D., NEUBERGER A. Carbohydrates in protein. 4. The determination of mannose in hen's-egg albumin by radioisotope dilution. Biochem J. 1962 May;83:335–341. doi: 10.1042/bj0830335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Foidart J. M., Bere E. W., Jr, Yaar M., Rennard S. I., Gullino M., Martin G. R., Katz S. I. Distribution and immunoelectron microscopic localization of laminin, a noncollagenous basement membrane glycoprotein. Lab Invest. 1980 Mar;42(3):336–342. [PubMed] [Google Scholar]
  9. Furthmayr H., Timpl R. Characterization of collagen peptides by sodium dodecylsulfate-polyacrylamide electrophoresis. Anal Biochem. 1971 Jun;41(2):510–516. doi: 10.1016/0003-2697(71)90173-4. [DOI] [PubMed] [Google Scholar]
  10. Glanville R. W., Rauter A., Fietzek P. P. Isolation and characterization of a native placental basement-membrane collagen and its component alpha chains. Eur J Biochem. 1979 Apr 2;95(2):383–389. doi: 10.1111/j.1432-1033.1979.tb12976.x. [DOI] [PubMed] [Google Scholar]
  11. Hahn E., Wick G., Pencev D., Timpl R. Distribution of basement membrane proteins in normal and fibrotic human liver: collagen type IV, laminin, and fibronectin. Gut. 1980 Jan;21(1):63–71. doi: 10.1136/gut.21.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Kresina T. F., Miller E. J. Isolation and characterization of basement membrane collagen from human placental tissue. Evidence for the presence of two genetically distinct collagen chains. Biochemistry. 1979 Jul 10;18(14):3089–3097. doi: 10.1021/bi00581a028. [DOI] [PubMed] [Google Scholar]
  14. Leivo I., Vaheri A., Timpl R., Wartiovaara J. Appearance and distribution of collagens and laminin in the early mouse embryo. Dev Biol. 1980 Apr;76(1):100–114. doi: 10.1016/0012-1606(80)90365-6. [DOI] [PubMed] [Google Scholar]
  15. Madri J. A., Roll F. J., Furthmayr H., Foidart J. M. Ultrastructural localization of fibronectin and laminin in the basement membranes of the murine kidney. J Cell Biol. 1980 Aug;86(2):682–687. doi: 10.1083/jcb.86.2.682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Müller H. A., Sinn M. Ein Beitrag zum dünnschichtchromatographischen Nachweis von Zuckern. Clin Chim Acta. 1977 Jul 1;78(1):17–21. doi: 10.1016/0009-8981(77)90333-3. [DOI] [PubMed] [Google Scholar]
  17. Risteli J., Bächinger H. P., Engel J., Furthmayr H., Timpl R. 7-S collagen: characterization of an unusual basement membrane structure. Eur J Biochem. 1980;108(1):239–250. doi: 10.1111/j.1432-1033.1980.tb04717.x. [DOI] [PubMed] [Google Scholar]
  18. Rohde H., Bächinger H. P., Timpl R. Characterization of pepsin fragments of laminin in a tumor basement membrane. Evidence for the existence of related proteins. Hoppe Seylers Z Physiol Chem. 1980 Nov;361(11):1651–1660. doi: 10.1515/bchm2.1980.361.2.1651. [DOI] [PubMed] [Google Scholar]
  19. Rohde H., Wick G., Timpl R. Immunochemical characterization of the basement membrane glycoprotein laminin. Eur J Biochem. 1979 Dec;102(1):195–201. doi: 10.1111/j.1432-1033.1979.tb06280.x. [DOI] [PubMed] [Google Scholar]
  20. Sage H., Woodbury R. G., Bornstein P. Structural studies on human type IV collagen. J Biol Chem. 1979 Oct 10;254(19):9893–9900. [PubMed] [Google Scholar]
  21. Shibata S., Nagasawa T., Miura K. Nephritogenoside, the receptor glycoprotein for concanavalin A in rat glomerular basement membrane. Demonstration of alpha-D-glucopyranosyl unit at the non-reducing terminus. Biochim Biophys Acta. 1977 Oct 25;499(3):392–403. doi: 10.1016/0304-4165(77)90070-8. [DOI] [PubMed] [Google Scholar]
  22. Timpl R., Rohde H., Robey P. G., Rennard S. I., Foidart J. M., Martin G. R. Laminin--a glycoprotein from basement membranes. J Biol Chem. 1979 Oct 10;254(19):9933–9937. [PubMed] [Google Scholar]
  23. 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]
  24. Wick G., Hönigsmann H., Timpl R. Immunofluorescence demonstration of type IV collagen and a noncollagenous glycoprotein in thickened vascular basal membranes in protoporphyria. J Invest Dermatol. 1979 Nov;73(5):335–338. doi: 10.1111/1523-1747.ep12550349. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES