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
. 1971 Jul;68(7):1450–1455. doi: 10.1073/pnas.68.7.1450

Antibodies Reactive with Native Lysozyme Elicited by a Completely Synthetic Antigen

Ruth Arnon 1, Elchanan Maron 1, Michael Sela 1, Christian B Anfinsen 1,*
PMCID: PMC389215  PMID: 5283934

Abstract

A synthetic peptide consisting of the aminoacid sequence of residues 64-82 of lysozyme, with alanine replacing cysteine as residue 76, was prepared by the solid-phase technique. Mild reduction followed by reoxidation in air of the deprotected peptide led to the formation of a closed loop containing an intrachain disulfide bond. A conjugate consisting of this “loop” attached to multi-poly(DL-alanyl)-poly(L-lysine) elicited, in rabbits and goats, the formation of antibodies capable of reacting with lysozyme and with the loop peptide prepared from it. These immunological interactions can be inhibited by either lysozyme or the loop peptide, but not by the performic acid-oxidized open-chain peptide. Thus, the antibodies elicited by the completely synthetic antigen show specificity toward the “loop” structure (residues 64-80) of native lysozyme.

Keywords: lysozyme residues 64-82, loop

Full text

PDF
1450

Images in this article

1453Image
on p.1453

Selected References

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

  1. ANFINSEN C. B., SELA M., COOKE J. P. The reversible reduction of disulfide bonds in polyalanyl ribonuclease. J Biol Chem. 1962 Jun;237:1825–1831. [PubMed] [Google Scholar]
  2. Arnon R., Sela M. Antibodies to a unique region in lysozyme provoked by a synthetic antigen conjugate. Proc Natl Acad Sci U S A. 1969 Jan;62(1):163–170. doi: 10.1073/pnas.62.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. BENJAMINI E., YOUNG J. D., SHIMIZU M., LEUNG C. Y. IMMUNOCHEMICAL STUDIES ON THE TOBACCO MOSAIC VIRUS PROTEIN. I. THE IMMUNOLOGICAL RELATIONSHIP OF THE TRYPTIC PEPTIDES OF TOBACCO MOSAIC VIRUS PROTEIN TO THE WHOLE PROTEIN. Biochemistry. 1964 Aug;3:1115–1120. doi: 10.1021/bi00896a018. [DOI] [PubMed] [Google Scholar]
  5. Brown J. R., Hartley B. S. Location of disulphide bridges by diagonal paper electrophoresis. The disulphide bridges of bovine chymotrypsinogen A. Biochem J. 1966 Oct;101(1):214–228. doi: 10.1042/bj1010214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  7. Fujio H., Imanishi M., Nishioka K., Amano T. Proof of independency of two antigenic sites in egg white lysozyme. Biken J. 1968 Sep;11(3):219–223. [PubMed] [Google Scholar]
  8. Haimovich J., Hurwitz E., Novik N., Sela M. Preparation of protein-bacteriophage conjugates and their use in detection of anti-protein antibodies. Biochim Biophys Acta. 1970 Apr 28;207(1):115–124. doi: 10.1016/0005-2795(70)90141-8. [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. Sanders M. M., Walsh K. A., Arnon R. Immunological cross-reaction between trypsin and chymotrypsin as a guide to structural homology. Biochemistry. 1970 May 26;9(11):2356–2363. doi: 10.1021/bi00813a020. [DOI] [PubMed] [Google Scholar]
  11. Schnabel E. Verbesserte Synthese von tert.-Butyloxycarbonyl-aminosäuren durch pH-Stat-Reaktion. Justus Liebigs Ann Chem. 1967;702:188–196. doi: 10.1002/jlac.19677020123. [DOI] [PubMed] [Google Scholar]
  12. Sela M. Antigenicity: some molecular aspects. Science. 1969 Dec 12;166(3911):1365–1374. doi: 10.1126/science.166.3911.1365. [DOI] [PubMed] [Google Scholar]
  13. Sela M. Immunological studies with synthetic polypeptides. Adv Immunol. 1966;5:29–129. doi: 10.1016/s0065-2776(08)60272-2. [DOI] [PubMed] [Google Scholar]
  14. Shinka S., Imanishi M., Miyagawa N., Amano T., Inoue M. Chemical studies on antigenic determinants of hen egg white lysozyme. I. Biken J. 1967 Jun;10(2):89–107. [PubMed] [Google Scholar]
  15. Ungar-Waron H., Hurwitz E., Jaton J. C., Sela M. Antibodies elicited with conjugates of nucleosides with synthetic polypeptides. Biochim Biophys Acta. 1967 May 30;138(3):513–531. doi: 10.1016/0005-2787(67)90548-5. [DOI] [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