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. 1998 Mar 16;17(6):1606–1613. doi: 10.1093/emboj/17.6.1606

Mac-2 binding protein is a cell-adhesive protein of the extracellular matrix which self-assembles into ring-like structures and binds beta1 integrins, collagens and fibronectin.

T Sasaki 1, C Brakebusch 1, J Engel 1, R Timpl 1
PMCID: PMC1170508  PMID: 9501082

Abstract

Human Mac-2 binding protein (M2BP) was prepared in recombinant form from the culture medium of 293 kidney cells and consisted of a 92 kDa subunit. The protein was obtained in a native state as indicated by CD spectroscopy, demonstrating alpha-helical and beta-type structure, and by protease resistance and immunological analysis. It was highly modified by N- and O-glycosylation but not by glycosaminoglycans. Ultracentrifugation showed non-covalent association into oligomers with molar masses of 1000-1500 kDa. Electron microscopy showed ring-like shapes with diameters of 30-40 nm. M2BP bound in solid-phase assays to collagens IV, V and VI, fibronectin and nidogen, but not to fibrillar collagens I and III or other basement membrane proteins. The protein also mediated adhesion of cell lines at comparable strength with laminin. Adhesion to M2BP was inhibited by antibodies to integrin beta1 subunits but not to alpha2 and alpha6 subunits, RGD peptide or lactose. This distinguishes cell adhesion of M2BP from that of laminin and excludes involvement of lactose-binding galectin-3. Immunological assays demonstrated variable secretion by cultured human cells of M2BP, which was detected in the extracellular matrix of several mouse tissues.

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Selected References

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

  1. Agrwal N., Sun Q., Wang S. Y., Wang J. L. Carbohydrate-binding protein 35. I. Properties of the recombinant polypeptide and the individuality of the domains. J Biol Chem. 1993 Jul 15;268(20):14932–14939. [PubMed] [Google Scholar]
  2. Aruffo A., Melnick M. B., Linsley P. S., Seed B. The lymphocyte glycoprotein CD6 contains a repeated domain structure characteristic of a new family of cell surface and secreted proteins. J Exp Med. 1991 Oct 1;174(4):949–952. doi: 10.1084/jem.174.4.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aumailley M., Mann K., von der Mark H., Timpl R. Cell attachment properties of collagen type VI and Arg-Gly-Asp dependent binding to its alpha 2(VI) and alpha 3(VI) chains. Exp Cell Res. 1989 Apr;181(2):463–474. doi: 10.1016/0014-4827(89)90103-1. [DOI] [PubMed] [Google Scholar]
  4. Aumailley M., Wiedemann H., Mann K., Timpl R. Binding of nidogen and the laminin-nidogen complex to basement membrane collagen type IV. Eur J Biochem. 1989 Sep 1;184(1):241–248. doi: 10.1111/j.1432-1033.1989.tb15013.x. [DOI] [PubMed] [Google Scholar]
  5. Battaglia C., Aumailley M., Mann K., Mayer U., Timpl R. Structural basis of beta 1 integrin-mediated cell adhesion to a large heparan sulfate proteoglycan from basement membranes. Eur J Cell Biol. 1993 Jun;61(1):92–99. [PubMed] [Google Scholar]
  6. Brakebusch C., Jallal B., Fusco O., Iacobelli S., Ullrich A. Expression of the 90K immunostimulator gene is controlled by a promoter with unique features. J Biol Chem. 1997 Feb 7;272(6):3674–3682. doi: 10.1074/jbc.272.6.3674. [DOI] [PubMed] [Google Scholar]
  7. Chicheportiche Y., Vassalli P. Cloning and expression of a mouse macrophage cDNA coding for a membrane glycoprotein of the scavenger receptor cysteine-rich domain family. J Biol Chem. 1994 Feb 25;269(8):5512–5517. [PubMed] [Google Scholar]
  8. Costell M., Mann K., Yamada Y., Timpl R. Characterization of recombinant perlecan domain I and its substitution by glycosaminoglycans and oligosaccharides. Eur J Biochem. 1997 Jan 15;243(1-2):115–121. doi: 10.1111/j.1432-1033.1997.t01-1-00115.x. [DOI] [PubMed] [Google Scholar]
  9. Dziadek M., Clements R., Mitrangas K., Reiter H., Fowler K. Analysis of degradation of the basement membrane protein nidogen, using a specific monoclonal antibody. Eur J Biochem. 1988 Feb 15;172(1):219–225. doi: 10.1111/j.1432-1033.1988.tb13876.x. [DOI] [PubMed] [Google Scholar]
  10. Engel J. Electron microscopy of extracellular matrix components. Methods Enzymol. 1994;245:469–488. doi: 10.1016/0076-6879(94)45024-2. [DOI] [PubMed] [Google Scholar]
  11. Fox J. W., Mayer U., Nischt R., Aumailley M., Reinhardt D., Wiedemann H., Mann K., Timpl R., Krieg T., Engel J. Recombinant nidogen consists of three globular domains and mediates binding of laminin to collagen type IV. EMBO J. 1991 Nov;10(11):3137–3146. doi: 10.1002/j.1460-2075.1991.tb04875.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Freeman M., Ashkenas J., Rees D. J., Kingsley D. M., Copeland N. G., Jenkins N. A., Krieger M. An ancient, highly conserved family of cysteine-rich protein domains revealed by cloning type I and type II murine macrophage scavenger receptors. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8810–8814. doi: 10.1073/pnas.87.22.8810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Friedman J., Trahey M., Weissman I. Cloning and characterization of cyclophilin C-associated protein: a candidate natural cellular ligand for cyclophilin C. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6815–6819. doi: 10.1073/pnas.90.14.6815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goldberger G., Bruns G. A., Rits M., Edge M. D., Kwiatkowski D. J. Human complement factor I: analysis of cDNA-derived primary structure and assignment of its gene to chromosome 4. J Biol Chem. 1987 Jul 25;262(21):10065–10071. [PubMed] [Google Scholar]
  15. Hall D. E., Reichardt L. F., Crowley E., Holley B., Moezzi H., Sonnenberg A., Damsky C. H. The alpha 1/beta 1 and alpha 6/beta 1 integrin heterodimers mediate cell attachment to distinct sites on laminin. J Cell Biol. 1990 Jun;110(6):2175–2184. doi: 10.1083/jcb.110.6.2175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Iacobelli S., Arnò E., D'Orazio A., Coletti G. Detection of antigens recognized by a novel monoclonal antibody in tissue and serum from patients with breast cancer. Cancer Res. 1986 Jun;46(6):3005–3010. [PubMed] [Google Scholar]
  17. Iacobelli S., Bucci I., D'Egidio M., Giuliani C., Natoli C., Tinari N., Rubistein M., Schlessinger J. Purification and characterization of a 90 kDa protein released from human tumors and tumor cell lines. FEBS Lett. 1993 Mar 15;319(1-2):59–65. doi: 10.1016/0014-5793(93)80037-u. [DOI] [PubMed] [Google Scholar]
  18. Iacobelli S., Sismondi P., Giai M., D'Egidio M., Tinari N., Amatetti C., Di Stefano P., Natoli C. Prognostic value of a novel circulating serum 90K antigen in breast cancer. Br J Cancer. 1994 Jan;69(1):172–176. doi: 10.1038/bjc.1994.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Inohara H., Akahani S., Koths K., Raz A. Interactions between galectin-3 and Mac-2-binding protein mediate cell-cell adhesion. Cancer Res. 1996 Oct 1;56(19):4530–4534. [PubMed] [Google Scholar]
  20. Inohara H., Raz A. Identification of human melanoma cellular and secreted ligands for galectin-3. Biochem Biophys Res Commun. 1994 Jun 30;201(3):1366–1375. doi: 10.1006/bbrc.1994.1854. [DOI] [PubMed] [Google Scholar]
  21. Jallal B., Powell J., Zachwieja J., Brakebusch C., Germain L., Jacobs J., Iacobelli S., Ullrich A. Suppression of tumor growth in vivo by local and systemic 90K level increase. Cancer Res. 1995 Aug 1;55(15):3223–3227. [PubMed] [Google Scholar]
  22. Jones N. H., Clabby M. L., Dialynas D. P., Huang H. J., Herzenberg L. A., Strominger J. L. Isolation of complementary DNA clones encoding the human lymphocyte glycoprotein T1/Leu-1. 1986 Sep 25-Oct 1Nature. 323(6086):346–349. doi: 10.1038/323346a0. [DOI] [PubMed] [Google Scholar]
  23. Kohfeldt E., Maurer P., Vannahme C., Timpl R. Properties of the extracellular calcium binding module of the proteoglycan testican. FEBS Lett. 1997 Sep 15;414(3):557–561. doi: 10.1016/s0014-5793(97)01070-3. [DOI] [PubMed] [Google Scholar]
  24. Koths K., Taylor E., Halenbeck R., Casipit C., Wang A. Cloning and characterization of a human Mac-2-binding protein, a new member of the superfamily defined by the macrophage scavenger receptor cysteine-rich domain. J Biol Chem. 1993 Jul 5;268(19):14245–14249. [PubMed] [Google Scholar]
  25. Linsley P. S., Horn D., Marquardt H., Brown J. P., Hellström I., Hellström K. E., Ochs V., Tolentino E. Identification of a novel serum protein secreted by lung carcinoma cells. Biochemistry. 1986 May 20;25(10):2978–2986. doi: 10.1021/bi00358a037. [DOI] [PubMed] [Google Scholar]
  26. Longo G., Natoli C., Rafanelli D., Tinari N., Morfini M., Rossi-Ferrini P., D'Ostilio N., Iacobelli S. Prognostic value of a novel circulating serum 90K antigen in HIV-infected haemophilia patients. Br J Haematol. 1993 Sep;85(1):207–209. doi: 10.1111/j.1365-2141.1993.tb08674.x. [DOI] [PubMed] [Google Scholar]
  27. Miller E. J., Rhodes R. K. Preparation and characterization of the different types of collagen. Methods Enzymol. 1982;82(Pt A):33–64. doi: 10.1016/0076-6879(82)82059-4. [DOI] [PubMed] [Google Scholar]
  28. Natali P. G., Wilson B. S., Imai K., Bigotti A., Ferrone S. Tissue distribution, molecular profile, and shedding of a cytoplasmic antigen identified by the monoclonal antibody 465.12S to human melanoma cells. Cancer Res. 1982 Feb;42(2):583–589. [PubMed] [Google Scholar]
  29. Nischt R., Pottgiesser J., Krieg T., Mayer U., Aumailley M., Timpl R. Recombinant expression and properties of the human calcium-binding extracellular matrix protein BM-40. Eur J Biochem. 1991 Sep 1;200(2):529–536. doi: 10.1111/j.1432-1033.1991.tb16214.x. [DOI] [PubMed] [Google Scholar]
  30. Odermatt E., Risteli J., van Delden V., Timpl R. Structural diversity and domain composition of a unique collagenous fragment (intima collagen) obtained from human placenta. Biochem J. 1983 May 1;211(2):295–302. doi: 10.1042/bj2110295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pan T. C., Sasaki T., Zhang R. Z., Fässler R., Timpl R., Chu M. L. Structure and expression of fibulin-2, a novel extracellular matrix protein with multiple EGF-like repeats and consensus motifs for calcium binding. J Cell Biol. 1993 Dec;123(5):1269–1277. doi: 10.1083/jcb.123.5.1269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Paulsson M., Aumailley M., Deutzmann R., Timpl R., Beck K., Engel J. Laminin-nidogen complex. Extraction with chelating agents and structural characterization. Eur J Biochem. 1987 Jul 1;166(1):11–19. doi: 10.1111/j.1432-1033.1987.tb13476.x. [DOI] [PubMed] [Google Scholar]
  33. Paulsson M., Yurchenco P. D., Ruben G. C., Engel J., Timpl R. Structure of low density heparan sulfate proteoglycan isolated from a mouse tumor basement membrane. J Mol Biol. 1987 Sep 20;197(2):297–313. doi: 10.1016/0022-2836(87)90125-2. [DOI] [PubMed] [Google Scholar]
  34. Rosenberg I., Cherayil B. J., Isselbacher K. J., Pillai S. Mac-2-binding glycoproteins. Putative ligands for a cytosolic beta-galactoside lectin. J Biol Chem. 1991 Oct 5;266(28):18731–18736. [PubMed] [Google Scholar]
  35. Sasaki T., Kostka G., Göhring W., Wiedemann H., Mann K., Chu M. L., Timpl R. Structural characterization of two variants of fibulin-1 that differ in nidogen affinity. J Mol Biol. 1995 Jan 20;245(3):241–250. doi: 10.1006/jmbi.1994.0020. [DOI] [PubMed] [Google Scholar]
  36. Sasaki T., Wiedemann H., Matzner M., Chu M. L., Timpl R. Expression of fibulin-2 by fibroblasts and deposition with fibronectin into a fibrillar matrix. J Cell Sci. 1996 Dec;109(Pt 12):2895–2904. doi: 10.1242/jcs.109.12.2895. [DOI] [PubMed] [Google Scholar]
  37. Taipale J., Keski-Oja J. Growth factors in the extracellular matrix. FASEB J. 1997 Jan;11(1):51–59. doi: 10.1096/fasebj.11.1.9034166. [DOI] [PubMed] [Google Scholar]
  38. Timpl R. Antibodies to collagens and procollagens. Methods Enzymol. 1982;82(Pt A):472–498. doi: 10.1016/0076-6879(82)82079-x. [DOI] [PubMed] [Google Scholar]
  39. Ullrich A., Sures I., D'Egidio M., Jallal B., Powell T. J., Herbst R., Dreps A., Azam M., Rubinstein M., Natoli C. The secreted tumor-associated antigen 90K is a potent immune stimulator. J Biol Chem. 1994 Jul 15;269(28):18401–18407. [PubMed] [Google Scholar]
  40. Vandenberg P., Kern A., Ries A., Luckenbill-Edds L., Mann K., Kühn K. Characterization of a type IV collagen major cell binding site with affinity to the alpha 1 beta 1 and the alpha 2 beta 1 integrins. J Cell Biol. 1991 Jun;113(6):1475–1483. doi: 10.1083/jcb.113.6.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Yu B., Wright S. D. LPS-dependent interaction of Mac-2-binding protein with immobilized CD14. J Inflamm. 1995;45(2):115–125. [PubMed] [Google Scholar]

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