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. 1974 May;139(2):359–368. doi: 10.1042/bj1390359

The interaction of α2-macroglobulin with proteinases. Binding and inhibition of mammalian collagenases and other metal proteinases

Zena Werb 1, Mary C Burleigh 1, Alan J Barrett 1, Phyllis M Starkey 1
PMCID: PMC1166291  PMID: 4374931

Abstract

1. Experiments were performed to determine whether the specific collagenases and other metal proteinases are bound and inhibited by α2-macroglobulin, as are endopeptidases of other classes. 2. A specific collagenase from rabbit synovial cells was inhibited by human serum. The inhibition could be attributed entirely to α2-macroglobulin; α1-trypsin inhibitor was not inhibitory. α2-Macroglobulin presaturated with trypsin or cathepsin B1 did not inhibit collagenase, and pretreatment of α2-macroglobulin with collagenase prevented subsequent reaction with trypsin. The binding of collagenase by α2-macroglobulin was not reversible in gel chromatography. 3. The collagenolytic activity of several rheumatoid synovial fluids was completely inhibited by incubation of the fluids with α2-macroglobulin. 4. The collagenase of human polymorphonuclear-leucocyte granules showed time-dependent inhibition by α2-macroglobulin. 5. The collagenolytic metal proteinase of Crotalus atrox venom was inhibited by α2-macroglobulin. 6. The collagenase of Clostridium histolyticum was bound by α2-macroglobulin, and inhibited more strongly with respect to collagen than with respect to a peptide substrate. 7. Thermolysin, the metal proteinase of Bacillus thermoproteolyticus, was bound and inhibited by α2-macroglobulin. 8. It was shown by polyacrylamidegel electrophoresis of reduced α2-macroglobulin in the presence of sodium dodecyl sulphate that synovial-cell collagenase, clostridial collagenase and thermolysin cleave the quarter subunit of α2-macroglobulin near its mid-point, as do serine proteinases. 9. The results are discussed in relation to previous work, and it is concluded that the characteristics of interaction of the metal proteinases with α2-macroglobulin are the same as those of other proteinases.

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Selected References

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

  1. Abe S., Nagai Y. Evidence for the presence of a complex of collagenase with alpha2-macroglobulin in human rheumatoid synovial fluid: a possible regulatory mechanism of collagenase activity in vivo. J Biochem. 1973 Apr;73(4):897–900. doi: 10.1093/oxfordjournals.jbchem.a130153. [DOI] [PubMed] [Google Scholar]
  2. Abe S., Nagai Y. Evidence for the presence of a latent form of collagenase in human rheumatoid synovial fluid. J Biochem. 1972 May;71(5):919–922. doi: 10.1093/oxfordjournals.jbchem.a129846. [DOI] [PubMed] [Google Scholar]
  3. Abe S., Nagai Y. Interaction between tadpole collagenase and human 2 -macroglobulin. Biochim Biophys Acta. 1972 Aug 31;278(1):125–132. doi: 10.1016/0005-2795(72)90113-4. [DOI] [PubMed] [Google Scholar]
  4. Barrett A. J. A new assay for cathepsin B1 and other thiol proteinases. Anal Biochem. 1972 May;47(1):280–293. doi: 10.1016/0003-2697(72)90302-8. [DOI] [PubMed] [Google Scholar]
  5. Barrett A. J. Human cathepsin B1. Purification and some properties of the enzyme. Biochem J. 1973 Apr;131(4):809–822. doi: 10.1042/bj1310809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Barrett A. J., Starkey P. M. The interaction of alpha 2-macroglobulin with proteinases. Characteristics and specificity of the reaction, and a hypothesis concerning its molecular mechanism. Biochem J. 1973 Aug;133(4):709–724. doi: 10.1042/bj1330709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bauer E. A., Eisen A. Z., Jeffrey J. J. Studies on purified rheumatoid synovial collagenase in vitro and in vivo. J Clin Invest. 1971 Oct;50(10):2056–2064. doi: 10.1172/JCI106699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chase T., Jr, Shaw E. p-Nitrophenyl-p'-guanidinobenzoate HCl: a new active site titrant for trypsin. Biochem Biophys Res Commun. 1967 Nov 30;29(4):508–514. doi: 10.1016/0006-291x(67)90513-x. [DOI] [PubMed] [Google Scholar]
  9. Colman P. M., Jansonius J. N., Matthews B. W. The structure of thermolysin: an electron density map at 2-3 A resolution. J Mol Biol. 1972 Oct 14;70(3):701–724. doi: 10.1016/0022-2836(72)90569-4. [DOI] [PubMed] [Google Scholar]
  10. Donoff R. B., McLennan J. E., Grillo H. C. Preparation and properties of collagenases from epithelium and mesenchyme of healing mammalian wounds. Biochim Biophys Acta. 1971 Mar 10;227(3):639–653. doi: 10.1016/0005-2744(71)90014-3. [DOI] [PubMed] [Google Scholar]
  11. Dresden M. H., Heilman S. A., Schmidt J. D. Collagenolytic enzymes in human neoplasms. Cancer Res. 1972 May;32(5):993–996. [PubMed] [Google Scholar]
  12. Eisen A. Z., Bauer E. A., Jeffrey J. J. Animal and human collagenases. J Invest Dermatol. 1970 Dec;55(6):359–373. doi: 10.1111/1523-1747.ep12260483. [DOI] [PubMed] [Google Scholar]
  13. Eisen A. Z., Bauer E. A., Jeffrey J. J. Human skin collagenase. The role of serum alpha-globulins in the control of activity in vivo and in vitro. Proc Natl Acad Sci U S A. 1971 Jan;68(1):248–251. doi: 10.1073/pnas.68.1.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Eisen A. Z., Bloch K. J., Sakai T. Inhibition of human skin collagenase by human serum. J Lab Clin Med. 1970 Feb;75(2):258–263. [PubMed] [Google Scholar]
  15. Evanson J. M., Jeffrey J. J., Krane S. M. Studies on collagenase from rheumatoid synovium in tissue culture. J Clin Invest. 1968 Dec;47(12):2639–2651. doi: 10.1172/JCI105947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fullmer H. M., Taylor R. E., Guthrie R. W. Human gingival collagenase: purification, molecular weight, and inhibitor studies. J Dent Res. 1972 Mar-Apr;51(2):349–355. doi: 10.1177/00220345720510022101. [DOI] [PubMed] [Google Scholar]
  17. Ganrot P. O. The combining ratio between trypsin and serum alpha-2-macroglobulin. Acta Chem Scand. 1966;20(8):2299–2300. doi: 10.3891/acta.chem.scand.20-2299. [DOI] [PubMed] [Google Scholar]
  18. HARTLEY B. S. Proteolytic enzymes. Annu Rev Biochem. 1960;29:45–72. doi: 10.1146/annurev.bi.29.070160.000401. [DOI] [PubMed] [Google Scholar]
  19. Harpel P. C. Studies on human plasma alpha 2-macroglobulin-enzyme interactions. Evidence for proteolytic modification of the subunit chain structure. J Exp Med. 1973 Sep 1;138(3):508–521. doi: 10.1084/jem.138.3.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Harris E. D., Jr, DiBona D. R., Krane S. M. Collagenases in human synovial fluid. J Clin Invest. 1969 Nov;48(11):2104–2113. doi: 10.1172/JCI106177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Harris E. D., Jr, Krane S. M. An endopeptidase from rheumatoid synovial tissue culture. Biochim Biophys Acta. 1972 Feb 28;258(2):566–576. doi: 10.1016/0005-2744(72)90249-5. [DOI] [PubMed] [Google Scholar]
  22. Jones J. M., Creeth J. M., Kekwick R. A. Thio reduction of human 2 -macroglobulin. The subunit structure. Biochem J. 1972 Mar;127(1):187–197. doi: 10.1042/bj1270187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lanchantin G. F., Plesset M. L., Friedmann J. A., Hart D. W. Dissociation of esterolytic and clotting activities of thrombin by trypsin-binding macroglobulin. Proc Soc Exp Biol Med. 1966 Feb;121(2):444–449. doi: 10.3181/00379727-121-30800. [DOI] [PubMed] [Google Scholar]
  24. Lazarus G. S., Daniels J. R., Brown R. S., Bladen H. A., Fullmer H. M. Degradation of collagen by a human granulocyte collagenolytic system. J Clin Invest. 1968 Dec;47(12):2622–2629. doi: 10.1172/JCI105945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lazarus G. S., Daniels J. R., Lian J., Burleigh M. C. Role of granulocyte collagenase in collagen degradation. Am J Pathol. 1972 Sep;68(3):565–578. [PMC free article] [PubMed] [Google Scholar]
  26. Mella K., Volz M., Pfleiderer G. Application of Crotalus atrox venom alpha-protease for amino acid sequence determination. Anal Biochem. 1967 Nov;21(2):219–226. doi: 10.1016/0003-2697(67)90183-2. [DOI] [PubMed] [Google Scholar]
  27. NORMAN P. S. Studies of the plasmin system. II. Inhibition of plasmin by serum or plasma. J Exp Med. 1958 Jul 1;108(1):53–68. doi: 10.1084/jem.108.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nagai Y., Lapiere C. M., Gross J. Tadpole collagenase. Preparation and purification. Biochemistry. 1966 Oct;5(10):3123–3130. doi: 10.1021/bi00874a007. [DOI] [PubMed] [Google Scholar]
  29. Peterkofsky B., Diegelmann R. Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. Biochemistry. 1971 Mar 16;10(6):988–994. doi: 10.1021/bi00782a009. [DOI] [PubMed] [Google Scholar]
  30. Reynolds J. M. Motivating office personnel for increased efficiency. Trans Eur Orthod Soc. 1973:379–385. [PubMed] [Google Scholar]
  31. Sakamoto S., Goldhaber P., Glimcher M. J. Further studies on the nature of the components in serum which inhibit mouse bone collagenase. Calcif Tissue Res. 1972;10(4):280–288. doi: 10.1007/BF02012559. [DOI] [PubMed] [Google Scholar]
  32. Sakamoto S., Goldhaber P., Glimcher M. J. Maintenance of mouse bone collagenase activity in the presence of serum protein by addition of trypsin. Proc Soc Exp Biol Med. 1972 Mar;139(3):1038–1041. doi: 10.3181/00379727-139-36293. [DOI] [PubMed] [Google Scholar]
  33. Starkey P. M., Barrett A. J. Inhibition by alpha-macroglobulin and other serum proteins. Biochem J. 1973 Apr;131(4):823–831. doi: 10.1042/bj1310823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  35. Werb Z., Burleigh M. C. A specific collagenase from rabbit fibroblasts in monolayer culture. Biochem J. 1974 Feb;137(2):373–385. doi: 10.1042/bj1370373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. de Vonne T. L., Mouray H., Berthillier G., Got R. Influence de l'alpha 2-macroglobuline du lapin sur l'activate enzymatique de la trypsine et de la chymotrypsine. Comp Biochem Physiol B. 1971 Oct;40(2):439–453. [PubMed] [Google Scholar]

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