Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1983 Mar 1;209(3):725–730. doi: 10.1042/bj2090725

Differential scanning calorimetry of alpha 2-macroglobulin and alpha 2-macroglobulin-proteinase complexes.

J F Chlebowski, K Williams
PMCID: PMC1154151  PMID: 6191752

Abstract

Differential scanning calorimetry is shown to detect substantial structural alterations occurring on the association of proteinases with the serum glycoprotein alpha 2-macroglobulin. At pH 7.5, the thermally induced unfolding of the macroglobulin occurs at approx. 60 degrees C with a transition enthalpy of 17 J/g. Association of active thermolysin, trypsin and papain shifts the transition temperature to 77 degrees C (transition enthalpy 5 J/g), indicating that a substantial conformational change accompanies the binding event. The stoicheiometry of the thermolysin--alpha 2-macroglobulin association producing this change appears to be unity, implying the presence of co-operative subunit interactions in the mechanism of association. The calorimetric method provides a novel approach for the evaluation of conformational variants induced on protein-protein association or pre-existing in the purified macroglobulin.

Full text

PDF
725

Selected References

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

  1. Barrett A. J., Brown M. A., Sayers C. A. The electrophoretically 'slow' and 'fast' forms of the alpha 2-macroglobulin molecule. Biochem J. 1979 Aug 1;181(2):401–418. doi: 10.1042/bj1810401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Biltonen R. L., Freire E. Thermodynamic characterization of conformational states of biological macromolecules using differential scanning calorimetry. CRC Crit Rev Biochem. 1978;5(2):85–124. doi: 10.3109/10409237809177141. [DOI] [PubMed] [Google Scholar]
  4. Chlebowski J. F., Mabrey S. Differential scanning calorimetry of apo-, apophosphoryl, and metalloalkaline phosphatases. J Biol Chem. 1977 Oct 25;252(20):7042–7052. [PubMed] [Google Scholar]
  5. Chlebowski J. F., Mabrey S., Falk M. C. Calorimetry of alkaline phosphatase. Stability of the monomer and effect of metal ion and phosphate binding on dimer stability. J Biol Chem. 1979 Jul 10;254(13):5745–5753. [PubMed] [Google Scholar]
  6. Dunn J. T., Spiro R. G. The alpha 2-macroglobulin of human plasma. I. Isolation and composition. J Biol Chem. 1967 Dec 10;242(23):5549–5555. [PubMed] [Google Scholar]
  7. Frénoy J. P., Bourrillon R., Lippoldt R., Edelhoch H. Stability and subunit structure of human alpha2-macroglobulin. J Biol Chem. 1977 Feb 25;252(4):1129–1133. [PubMed] [Google Scholar]
  8. Frénoy J. P., Bourrillon R. Studies on the structure of human alpha2-macroglobulin. IV. Analysis of the microheterogeneity by isoelectric focusing. Biochim Biophys Acta. 1974 Nov 5;371(1):168–176. doi: 10.1016/0005-2795(74)90166-4. [DOI] [PubMed] [Google Scholar]
  9. Howard J. B. Methylamine reaction and denaturation-dependent fragmentation of complement component 3. Comparison with alpha2-macroglobulin. J Biol Chem. 1980 Aug 10;255(15):7082–7084. [PubMed] [Google Scholar]
  10. Howard J. B. Reactive site in human alpha 2-macroglobulin: circumstantial evidence for a thiolester. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2235–2239. doi: 10.1073/pnas.78.4.2235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Privalov P. L., Khechinashvili N. N. A thermodynamic approach to the problem of stabilization of globular protein structure: a calorimetric study. J Mol Biol. 1974 Jul 5;86(3):665–684. doi: 10.1016/0022-2836(74)90188-0. [DOI] [PubMed] [Google Scholar]
  13. Privalov P. L. Stability of proteins: small globular proteins. Adv Protein Chem. 1979;33:167–241. doi: 10.1016/s0065-3233(08)60460-x. [DOI] [PubMed] [Google Scholar]
  14. Sottrup-Jensen L., Petersen T. E., Magnusson S. Trypsin-induced activation of the thiol esters in alpha 2-macroglobulin generates a short-lived intermediate ('nascent' alpha 2-M) that can react rapidly to incorporate not only methylamine or putrescine but also proteins lacking proteinase activity. FEBS Lett. 1981 Jun 1;128(1):123–126. doi: 10.1016/0014-5793(81)81096-4. [DOI] [PubMed] [Google Scholar]
  15. Swenson R. P., Howard J. B. Characterization of alkylamine-sensitive site in alpha 2-macroglobulin. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4313–4316. doi: 10.1073/pnas.76.9.4313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Swenson R. P., Howard J. B. Structural characterization of human alpha2-macroglobulin subunits. J Biol Chem. 1979 Jun 10;254(11):4452–4456. [PubMed] [Google Scholar]
  17. Takahashi K., Sturtevant J. M. Thermal denaturation of streptomyces subtilisin inhibitor, subtilisin BPN', and the inhibitor-subtilisin complex. Biochemistry. 1981 Oct 13;20(21):6185–6190. doi: 10.1021/bi00524a042. [DOI] [PubMed] [Google Scholar]
  18. Topping R. M., Craven A. H. The identification of distinctive forms of human alpha 2-macroglobulin by using the numerical relationship between trypsin binding in alpha- and beta-modes. Biochem J. 1979 Feb 1;177(2):501–508. doi: 10.1042/bj1770501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Topping R. M., Seilman S. A four-straight-line model for the proteinase-binding characteristics of human blood serum. Biochem J. 1979 Feb 1;177(2):493–499. doi: 10.1042/bj1770493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vickers L. P., Donovan J. W., Schachman H. K. Differential scanning calorimetry of asparate transcarbamoylase and its isolate subunits. J Biol Chem. 1978 Dec 10;253(23):8493–8498. [PubMed] [Google Scholar]
  21. Werb Z., Burleigh M. C., Barrett A. J., Starkey P. M. The interaction of alpha2-macroglobulin with proteinases. Binding and inhibition of mammalian collagenases and other metal proteinases. Biochem J. 1974 May;139(2):359–368. doi: 10.1042/bj1390359. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES