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. 1985 Jul;4(7):1731–1737. doi: 10.1002/j.1460-2075.1985.tb03843.x

Towards assignment of secondary structures by anti-peptide antibodies. Specificity of the immune response to a beta-turn.

U Schulze-Gahmen, H Prinz, U Glatter, K Beyreuther
PMCID: PMC554410  PMID: 2411545

Abstract

In an attempt to assign secondary structure elements to protein primary structures with antibodies, we synthesized a model peptide (beta-peptide: TVTVTDPGQTVTY) with a putative beta-turn structure and analysed the anti-peptide antibodies for their specificity towards the turn sequence. At least 50% of the peptide fraction adopts the intended conformation of a beta-turn (DPGQ) inserted between the two segments of an antiparallel beta-sheet structure. The specific anti-beta-peptide antibodies of the hyperimmune response bind the beta-turn containing epitope of the immunogenic beta-peptide with a three orders of magnitude higher affinity than the synthetic control peptide (Gly-peptide: GGGGGDPGQGGGG). The affinity of the antibodies with specificity for the beta-turn region increases from the primary to the hyperimmune response. Therefore, probing of secondary structure elements, i.e., of individual beta-turn regions, by anti-peptide antibodies now seems feasible for proteins of known sequence and may result in sequence assignments of secondary structures.

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Selected References

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  1. Antoine J. C., Avrameas S. Correlations between immunoglobulin- and antibody-synthesizing cells during primary and secondary immune responses of rats immunized with peroxidase. Immunology. 1976 Apr;30(4):537–547. [PMC free article] [PubMed] [Google Scholar]
  2. Berzofsky J. A., Schechter A. N. The concepts of crossreactivity and specificity in immunology. Mol Immunol. 1981 Aug;18(8):751–763. doi: 10.1016/0161-5890(81)90067-5. [DOI] [PubMed] [Google Scholar]
  3. Beyreuther K., Raufuss H., Schrecker O., Hengstenberg W. The phosphoenolpyruvate-dependent phosphotransferase system of Staphylococcus aureus. 1. Amino-acid sequence of the phosphocarrier protein HPr. Eur J Biochem. 1977 May 2;75(1):275–286. doi: 10.1111/j.1432-1033.1977.tb11527.x. [DOI] [PubMed] [Google Scholar]
  4. Brack A., Orgel L. E. Beta structures of alternating polypeptides and their possible prebiotic significance. Nature. 1975 Jul 31;256(5516):383–387. doi: 10.1038/256383a0. [DOI] [PubMed] [Google Scholar]
  5. Brahms S., Brahms J. Determination of protein secondary structure in solution by vacuum ultraviolet circular dichroism. J Mol Biol. 1980 Apr;138(2):149–178. doi: 10.1016/0022-2836(80)90282-x. [DOI] [PubMed] [Google Scholar]
  6. Brahms S., Brahms J., Spach G., Brack A. Identification of beta,beta-turns and unordered conformations in polypeptide chains by vacuum ultraviolet circular dichroism. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3208–3212. doi: 10.1073/pnas.74.8.3208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chang C. T., Wu C. S., Yang J. T. Circular dichroic analysis of protein conformation: inclusion of the beta-turns. Anal Biochem. 1978 Nov;91(1):13–31. doi: 10.1016/0003-2697(78)90812-6. [DOI] [PubMed] [Google Scholar]
  8. Chou P. Y., Fasman G. D. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. doi: 10.1002/9780470122921.ch2. [DOI] [PubMed] [Google Scholar]
  9. Green N., Alexander H., Olson A., Alexander S., Shinnick T. M., Sutcliffe J. G., Lerner R. A. Immunogenic structure of the influenza virus hemagglutinin. Cell. 1982 Mar;28(3):477–487. doi: 10.1016/0092-8674(82)90202-1. [DOI] [PubMed] [Google Scholar]
  10. Gutte B., Lin M. C., Caldi D. G., Merrifield R. B. Reactivation of des(119-, 120-, or 121-124) ribonuclease A by mixture with synthetic COOH-terminal peptides of varying lengths. J Biol Chem. 1972 Aug 10;247(15):4763–4767. [PubMed] [Google Scholar]
  11. Gutte B., Merrifield R. B. The synthesis of ribonuclease A. J Biol Chem. 1971 Mar 25;246(6):1922–1941. [PubMed] [Google Scholar]
  12. Kullmann W. Design, synthesis, and binding characteristics of an opiate receptor mimetic peptide. J Med Chem. 1984 Feb;27(2):106–115. doi: 10.1021/jm00368a002. [DOI] [PubMed] [Google Scholar]
  13. Lewis P. N., Momany F. A., Scheraga H. A. Folding of polypeptide chains in proteins: a proposed mechanism for folding. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2293–2297. doi: 10.1073/pnas.68.9.2293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. MALLEY A., SAHA A., HALLIDAY W. J. IMMUNOCHEMICAL STUDIES OF HEMOCYANIN FROM THE GIANT KEYHOLE LIMPET (MEGATHURA CRENULATA) AND THE HORSESHOE CRAB (LIMULUS POLYHEMUS). J Immunol. 1965 Jul;95:141–147. [PubMed] [Google Scholar]
  15. Offord R. E. Electrophoretic mobilities of peptides on paper and their use in the determination of amide groups. Nature. 1966 Aug 6;211(5049):591–593. doi: 10.1038/211591a0. [DOI] [PubMed] [Google Scholar]
  16. Pfaff E., Mussgay M., Böhm H. O., Schulz G. E., Schaller H. Antibodies against a preselected peptide recognize and neutralize foot and mouth disease virus. EMBO J. 1982;1(7):869–874. doi: 10.1002/j.1460-2075.1982.tb01262.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Poljak R. J., Amzel L. M., Chen B. L., Chiu Y. Y., Phizackerley R. P., Saul F., Ysern X. Three-dimensional structure and diversity of immunoglobulins. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 2):639–645. doi: 10.1101/sqb.1977.041.01.073. [DOI] [PubMed] [Google Scholar]
  18. Rose G. D., Wetlaufer D. B. The number of turns in globular proteins. Nature. 1977 Aug 25;268(5622):769–770. doi: 10.1038/268769a0. [DOI] [PubMed] [Google Scholar]
  19. Sachs D. H., Schechter A. N., Eastlake A., Anfinsen C. B. An immunologic approach to the conformational equilibria of polypeptides. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3790–3794. doi: 10.1073/pnas.69.12.3790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tarr G. E., Beecher J. F., Bell M., McKean D. J. Polyquarternary amines prevent peptide loss from sequenators. Anal Biochem. 1978 Feb;84(2):622–7?0=ENG. doi: 10.1016/0003-2697(78)90086-6. [DOI] [PubMed] [Google Scholar]
  21. Wagner G., Wüthrich K. Sequential resonance assignments in protein 1H nuclear magnetic resonance spectra. Basic pancreatic trypsin inhibitor. J Mol Biol. 1982 Mar 5;155(3):347–366. doi: 10.1016/0022-2836(82)90009-2. [DOI] [PubMed] [Google Scholar]
  22. Walter G., Scheidtmann K. H., Carbone A., Laudano A. P., Doolittle R. F. Antibodies specific for the carboxy- and amino-terminal regions of simian virus 40 large tumor antigen. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5197–5200. doi: 10.1073/pnas.77.9.5197. [DOI] [PMC free article] [PubMed] [Google Scholar]

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