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. 1984 Jun 1;98(6):1937–1946. doi: 10.1083/jcb.98.6.1937

Chick myotendinous antigen. II. A novel extracellular glycoprotein complex consisting of large disulfide-linked subunits

PMCID: PMC2113072  PMID: 6202699

Abstract

This report describes the biochemical characterization of a novel extracellular matrix component, " myotendinous antigen," which appears early in chick limb morphogenesis at sites connecting developing muscle fibers, tendons, and bone ( Chiquet , M., and D. Fambrough , 1984; J. Cell Biol., 98:1926-1936). This extracellular matrix antigen is a major component of the secretory proteins released into the medium by fibroblast and muscle cultures; the soluble form is characterized here. This form of myotendinous antigen is a large glycoprotein complex consisting of several disulfide linked subunits (Mr approximately 150,000-240,000). The differently sized antigen subunits are related, since they yielded very similar proteolytic cleavage patterns. M1 antibody can bind to the denatured subunits. The antigen subunits, as well as a Mr approximately 80,000 pepsin-resistant antigenic domain derived from them, are resistant to bacterial collagenase. Despite possessing subunits similar in size to fibronectin, myotendinous antigen appears to be both structurally and antigenically unrelated to fibronectin or to other known extracellular matrix components. About seven times more M1 antigen per cell nucleus was released into the medium in fibroblast as compared to muscle cultures. In muscle conditioned medium, myotendinous antigen is noncovalently complexed to very high molecular weight material that could be heavily labeled by [3H]glucosamine and [35S]sulfate. This material is sensitive to chondroitinase ABC and hence appears to contain sulfated glycosaminoglycans. We speculate that myotendinous antigen might interact with proteoglycans on the surface of muscle fibers, thereby acting as a link to tendons.

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Selected References

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  1. Alexander S. S., Jr, Colonna G., Edelhoch H. The structure and stability of human plasma cold-insoluble globulin. J Biol Chem. 1979 Mar 10;254(5):1501–1505. [PubMed] [Google Scholar]
  2. Anderson M. J., Fambrough D. M. Aggregates of acetylcholine receptors are associated with plaques of a basal lamina heparan sulfate proteoglycan on the surface of skeletal muscle fibers. J Cell Biol. 1983 Nov;97(5 Pt 1):1396–1411. doi: 10.1083/jcb.97.5.1396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bornstein P., Sage H. Structurally distinct collagen types. Annu Rev Biochem. 1980;49:957–1003. doi: 10.1146/annurev.bi.49.070180.004521. [DOI] [PubMed] [Google Scholar]
  4. Bornstein P. The biosynthesis of collagen. Annu Rev Biochem. 1974;43(0):567–603. doi: 10.1146/annurev.bi.43.070174.003031. [DOI] [PubMed] [Google Scholar]
  5. Bächinger H. P., Fessler L. I., Fessler J. H. Mouse procollagen IV. Characterization and supramolecular association. J Biol Chem. 1982 Aug 25;257(16):9796–9803. [PubMed] [Google Scholar]
  6. Caravatti M., Perriard J. C., Eppenberger H. M. Developmental regulation of creatine kinase isoenzymes in myogenic cell cultures from chicken. Biosynthesis of creatine kinase subunits M and B. J Biol Chem. 1979 Feb 25;254(4):1388–1394. [PubMed] [Google Scholar]
  7. Carlin B., Jaffe R., Bender B., Chung A. E. Entactin, a novel basal lamina-associated sulfated glycoprotein. J Biol Chem. 1981 May 25;256(10):5209–5214. [PubMed] [Google Scholar]
  8. Carrino D. A., Caplan A. I. Isolation and preliminary characterization of proteoglycans synthesized by skeletal muscle. J Biol Chem. 1982 Dec 10;257(23):14145–14154. [PubMed] [Google Scholar]
  9. Chen L. B., Murray A., Segal R. A., Bushnell A., Walsh M. L. Studies on intercellular LETS glycoprotein matrices. Cell. 1978 Jun;14(2):377–391. doi: 10.1016/0092-8674(78)90123-x. [DOI] [PubMed] [Google Scholar]
  10. Chiquet M., Fambrough D. M. Chick myotendinous antigen. I. A monoclonal antibody as a marker for tendon and muscle morphogenesis. J Cell Biol. 1984 Jun;98(6):1926–1936. doi: 10.1083/jcb.98.6.1926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chiquet M., Puri E. C., Turner D. C. Fibronectin mediates attachment of chicken myoblasts to a gelatin-coated substratum. J Biol Chem. 1979 Jun 25;254(12):5475–5482. [PubMed] [Google Scholar]
  12. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  13. Ehrismann R., Chiquet M., Turner D. C. Mode of action of fibronectin in promoting chicken myoblast attachment. Mr = 60,000 gelatin-binding fragment binds native fibronectin. J Biol Chem. 1981 Apr 25;256(8):4056–4062. [PubMed] [Google Scholar]
  14. Engvall E., Bell M. L., Carlsson R. N., Miller E. J., Ruoslahti E. Nonhelical, fibronectin-binding basement-membrane collagen from endodermal cell culture. Cell. 1982 Jun;29(2):475–482. doi: 10.1016/0092-8674(82)90164-7. [DOI] [PubMed] [Google Scholar]
  15. Eyre D. R. Collagen: molecular diversity in the body's protein scaffold. Science. 1980 Mar 21;207(4437):1315–1322. doi: 10.1126/science.7355290. [DOI] [PubMed] [Google Scholar]
  16. Fessler L. I., Fessler J. H. Identification of the carboxyl peptides of mouse procollagen IV and its implications for the assembly and structure of basement membrane procollagen. J Biol Chem. 1982 Aug 25;257(16):9804–9810. [PubMed] [Google Scholar]
  17. Fessler L. I., Kumamoto C. A., Meis M. E., Fessler J. H. Assembly and processing of procollagen V (AB) in chick blood vessels and other tissues. J Biol Chem. 1981 Sep 25;256(18):9640–9645. [PubMed] [Google Scholar]
  18. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gardner J. M., Fambrough D. M. Fibronectin expression during myogenesis. J Cell Biol. 1983 Feb;96(2):474–485. doi: 10.1083/jcb.96.2.474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hewitt A. T., Varner H. H., Silver M. H., Dessau W., Wilkes C. M., Martin G. R. The isolation and partial characterization of chondronectin, an attachment factor for chondrocytes. J Biol Chem. 1982 Mar 10;257(5):2330–2334. [PubMed] [Google Scholar]
  21. Hogan B. L., Taylor A., Kurkinen M., Couchman J. R. Synthesis and localization of two sulphated glycoproteins associated with basement membranes and the extracellular matrix. J Cell Biol. 1982 Oct;95(1):197–204. doi: 10.1083/jcb.95.1.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hynes R. O., Martin G. S., Shearer M., Critchley D. R., Epstein C. J. Viral transformation of rat myoblasts: effects on fusion and surface properties. Dev Biol. 1976 Jan;48(1):35–46. doi: 10.1016/0012-1606(76)90043-9. [DOI] [PubMed] [Google Scholar]
  23. Hynes R. O., Yamada K. M. Fibronectins: multifunctional modular glycoproteins. J Cell Biol. 1982 Nov;95(2 Pt 1):369–377. doi: 10.1083/jcb.95.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kewley M. A., Steven F. S., Williams G. Preparation of a specific antiserum towards the microfibrillar protein of elastic tissues. Immunology. 1977 Apr;32(4):483–489. [PMC free article] [PubMed] [Google Scholar]
  25. Krieg T., Timpl R., Alitalo K., Kurkinen M., Vaheri A. Type III procollagen is the major collageneous component produced by a continuous rhabdomyosarcoma cell line. FEBS Lett. 1979 Aug 15;104(2):405–409. doi: 10.1016/0014-5793(79)80863-7. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  28. Mayer B. W., Jr, Hay E. D., Hynes R. O. Immunocytochemical localization of fibronectin in embryonic chick trunk and area vasculosa. Dev Biol. 1981 Mar;82(2):267–286. doi: 10.1016/0012-1606(81)90451-6. [DOI] [PubMed] [Google Scholar]
  29. Millis A. J., Hoyle M. Fibroblast-conditioned medium contains cell surface proteins required for cell attachment and spreading. Nature. 1978 Feb 16;271(5646):668–669. doi: 10.1038/271668a0. [DOI] [PubMed] [Google Scholar]
  30. Muir L. W., Bornstein P., Ross R. A presumptive subunit of elastic fiber microfibrils secreted by arterial smooth-muscle cells in culture. Eur J Biochem. 1976 Apr 15;64(1):105–114. doi: 10.1111/j.1432-1033.1976.tb10278.x. [DOI] [PubMed] [Google Scholar]
  31. Neff N. T., Lowrey C., Decker C., Tovar A., Damsky C., Buck C., Horwitz A. F. A monoclonal antibody detaches embryonic skeletal muscle from extracellular matrices. J Cell Biol. 1982 Nov;95(2 Pt 1):654–666. doi: 10.1083/jcb.95.2.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Raugi G. J., Mumby S. M., Abbott-Brown D., Bornstein P. Thrombospondin: synthesis and secretion by cells in culture. J Cell Biol. 1982 Oct;95(1):351–354. doi: 10.1083/jcb.95.1.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Reese C. A., Mayne R. Minor collagens of chicken hyaline cartilage. Biochemistry. 1981 Sep 15;20(19):5443–5448. doi: 10.1021/bi00522a014. [DOI] [PubMed] [Google Scholar]
  34. Ross R., Bornstein P. The elastic fiber. I. The separation and partial characterization of its macromolecular components. J Cell Biol. 1969 Feb;40(2):366–381. doi: 10.1083/jcb.40.2.366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sasse J., von der Mark H., Kühl U., Dessau W., von der Mark K. Origin of collagen types I, III, and V in cultures of avian skeletal muscle. Dev Biol. 1981 Apr 15;83(1):79–89. doi: 10.1016/s0012-1606(81)80010-3. [DOI] [PubMed] [Google Scholar]
  36. Schmid T. M., Conrad H. E. A unique low molecular weight collagen secreted by cultured chick embryo chondrocytes. J Biol Chem. 1982 Oct 25;257(20):12444–12450. [PubMed] [Google Scholar]
  37. Sear C. H., Kewley M. A., Jones C. J., Grant M. E., Jackson D. S. The identification of glycoproteins associated with elastic-tissue microfibrils. Biochem J. 1978 Mar 15;170(3):715–718. doi: 10.1042/bj1700715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Trotter J. A., Corbett K., Avner B. P. Structure and function of the murine muscle-tendon junction. Anat Rec. 1981 Oct;201(2):293–302. doi: 10.1002/ar.1092010209. [DOI] [PubMed] [Google Scholar]
  41. Turner D. C. Differentiation in cultures derived from embryonic chicken muscle: the postmitotic, fusion-capable myoblast as a distinct cell type. Differentiation. 1978 Mar 13;10(2):81–93. doi: 10.1111/j.1432-0436.1978.tb00949.x. [DOI] [PubMed] [Google Scholar]
  42. Turner D. C., Maier V., Eppenberger H. M. Creatine kinase and aldolase isoenzyme transitions in cultures of chick skeletal muscle cells. Dev Biol. 1974 Mar;37(1):63–89. doi: 10.1016/0012-1606(74)90170-5. [DOI] [PubMed] [Google Scholar]
  43. Yamada K. M., Yamada S. S., Pastan I. Cell surface protein partially restores morphology, adhesiveness, and contact inhibition of movement to transformed fibroblasts. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1217–1221. doi: 10.1073/pnas.73.4.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. von der Mark H., von der Mark H. Isolation and characterization of collagen A and B chains from chick embryos. FEBS Lett. 1979 Mar 1;99(1):101–105. doi: 10.1016/0014-5793(79)80258-6. [DOI] [PubMed] [Google Scholar]

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