Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1972 Dec;69(12):3790–3794. doi: 10.1073/pnas.69.12.3790

An Immunologic Approach to the Conformational Equilibria of Polypeptides

David H Sachs 1, Alan N Schechter 1, Ann Eastlake 1, Christian B Anfinsen 1
PMCID: PMC389874  PMID: 4118873

Abstract

Antibodies reactive with distinct regions of the staphylococcal nuclease molecule were prepared both by immunization with polypeptide fragments of nuclease and by immunization with intact nuclease followed by fractionation of the antiserum on immuno-absorbent columns bearing the corresponding fragments. Comparisons of the interactions of these antibody preparations with nuclease, by quantitative precipitin assays and enzyme inhibition studies, showed marked differences attributable to the conformation of the immunizing antigens. An interpretive model is proposed in which antibodies fractionated from anti-nuclease serum react effectively only with the “native format determinants” of polypeptide fragments of nuclease. It is postulated that such determinants are generated in polypeptide fragments by spontaneous and reversible folding of the polypeptide chain. The model permits experimental determination of parameters, Kconf, for the proposed conformational equilibria.

Full text

PDF
3790

Selected References

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

  1. Arnon R. A selective fractionation of anti-lysozyme antibodies of different determinant specificities. Eur J Biochem. 1968 Sep 24;5(4):583–589. doi: 10.1111/j.1432-1033.1968.tb00409.x. [DOI] [PubMed] [Google Scholar]
  2. Atassi M. Z., Saplin B. J. Immunochemistry of sperm whale myoglobin. I. The specific interaction of some tryptic peptides and of peptides containing all the reactive regions of the antigen. Biochemistry. 1968 Feb;7(2):688–698. doi: 10.1021/bi00842a026. [DOI] [PubMed] [Google Scholar]
  3. Crumpton M. J., Wilkinson J. M. The immunological activity of some of the chymotryptic peptides of sperm-whale myoglobin. Biochem J. 1965 Mar;94(3):545–556. doi: 10.1042/bj0940545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cuatrecasas P., Fuchs S., Anfinsen C. B. Catalytic properties and specificity of the extracellular nuclease of Staphylococcus aureus. J Biol Chem. 1967 Apr 10;242(7):1541–1547. [PubMed] [Google Scholar]
  5. Cuatrecasas P., Wilchek M., Anfinsen C. B. Selective enzyme purification by affinity chromatography. Proc Natl Acad Sci U S A. 1968 Oct;61(2):636–643. doi: 10.1073/pnas.61.2.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fujio H., Sakato N., Amano T. The immunological properties of region specific antibodies directed to hen egg-white lysozyme. Biken J. 1971 Dec;14(4):395–404. [PubMed] [Google Scholar]
  7. Hamlin J., Zabin I. -Galactosidase: immunological activity of ribosome-bound, growing polypeptide chains. Proc Natl Acad Sci U S A. 1972 Feb;69(2):412–416. doi: 10.1073/pnas.69.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Isagholian L. B., Brown R. K. Interaction of a peptide with antibody to oxidized ribonuclease. Immunochemistry. 1970 Feb;7(2):167–174. doi: 10.1016/0019-2791(70)90152-7. [DOI] [PubMed] [Google Scholar]
  9. Merrifield R. B. Automated synthesis of peptides. Science. 1965 Oct 8;150(3693):178–185. doi: 10.1126/science.150.3693.178. [DOI] [PubMed] [Google Scholar]
  10. Morávek L., Anfinsen C. B., Cone J. L., Taniuchi H. The large scale preparation of an extracellular nuclease of Staphylococcus aureus. J Biol Chem. 1969 Jan 25;244(2):497–499. [PubMed] [Google Scholar]
  11. Omenn G. S., Ontjes D. A., Anfinsen C. B. Immunochemistry of staphylococcal nuclease. II. Inhibition and binding studies with sequence fragments. Biochemistry. 1970 Jan 20;9(2):313–321. doi: 10.1021/bi00804a018. [DOI] [PubMed] [Google Scholar]
  12. Sachs D. H., Schechter A. N., Eastlake A., Anfinsen C. B. Antibodies to a distinct antigenic determinant of staphylococcal nuclease. J Immunol. 1972 Dec;109(6):1300–1310. [PubMed] [Google Scholar]
  13. Sachs D. H., Schechter A. N., Eastlake A., Anfinsen C. B. Inactivation of staphylococcal nuclease by the binding of antibodies to a distinct antigenic determinant. Biochemistry. 1972 Nov 7;11(23):4268–4273. doi: 10.1021/bi00773a012. [DOI] [PubMed] [Google Scholar]
  14. Schechter A. N., Morávek L., Anfinsen C. B. Suppression of hydrogen exchange in staphylococcal nuclease by ligands. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1478–1485. doi: 10.1073/pnas.61.4.1478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schechter B., Conway-Jacobs A., Sela M. Conformational changes in a synthetic antigen induced by specific antibodies. Eur J Biochem. 1971 Jun 11;20(3):321–324. doi: 10.1111/j.1432-1033.1971.tb01396.x. [DOI] [PubMed] [Google Scholar]
  16. Taniuchi H., Anfinsen C. B. An experimental approach to the study of the folding of staphylococcal nuclease. J Biol Chem. 1969 Jul 25;244(14):3864–3875. [PubMed] [Google Scholar]
  17. Taniuchi H., Anfinsen C. B. Simultaneous formation of two alternative enzymology active structures by complementation of two overlapping fragments of staphylococcal nuclease. J Biol Chem. 1971 Apr 10;246(7):2291–2301. [PubMed] [Google Scholar]
  18. Taniuchi H., Anfinsen C. B., Sodja A. Nuclease-T: an active derivative of staphylococcal nuclease composed of two noncovalently bonded peptide fragments. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1235–1242. doi: 10.1073/pnas.58.3.1235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Taniuchi H., Anfinsen C. B. The amino acid sequence of an extracellular nuclease of Staphylococcus aureus. I. Linear order of the fragments produced by cleavage with cyanogen bromide. J Biol Chem. 1966 Oct 10;241(19):4366–4385. [PubMed] [Google Scholar]
  20. Taniuchi H., Morávek L., Anfinsen C. B. Ligand-induced resistance of staphylococcal nuclease and nuclease-T to proteolysis by subtilisin, alpha-chymotrypsin, and thermolysin. J Biol Chem. 1969 Sep 10;244(17):4600–4606. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES