Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1970 Sep 1;132(3):385–400. doi: 10.1084/jem.132.3.385

PHYLOGENY OF IMMUNOGLOBULIN STRUCTURE AND FUNCTION

V. VALENCES AND ASSOCIATION CONSTANTS OF TELEOST ANTIBODIES TO A HAPTENIC DETERMINANT

L W Clem 1, P A Small Jr 1
PMCID: PMC2138810  PMID: 4109109

Abstract

The giant grouper, Epinephelus itaira, was shown to synthesize 16 and 6.4S antibodies specific for the dinitrophenyl determinant (DNP). Sera obtained at various intervals between 1 month and 2 yr after initial immunization contained both species of antibody; no temporal synthesis was evident. Equilibrium dialysis studies employing ε-dinitrophenyl-amino caproic acid were conducted with purified grouper antibodies specific for DNP. The 16S antibody preparations obtained at 1 and 2 months of immunization showed heterogeneity of hapten binding indicative of two populations of combining sites. One-half of these sites (an average of four sites per 16S molecule) exhibited an average intrinsic association constant (Ko) of ∼106 M –1; the Ko of the remaining four-sites was ∼104 M –1. Thus, the valence of the grouper 16S antibody molecule appears to be eight although the distribution of the high and low Ko sites is unknown, i.e., are they each on the same or on different molecules? The 16S antibody preparations obtained after more prolonged immunization exhibited increasingly lower Ko values; the so-called low Ko sites were no longer detectable. These findings are in contrast to reports of rabbit IgG antibodies showing an increase in Ko with increased time. The 6.4S antibody preparations obtained from the 1 and 2 month antisera had Ko values of ∼106 M –1 and a valence of one. These antibodies would not precipitate with antigen. The 6.4S antibody preparations obtained at later times showed decreasing Ko values comparable to those of the 16S antibodies from the same bleedings. Studies on the thermodynamic parameters of the hapten-antibody interaction showed the grouper 16 and 6.4S antibodies to be similar to each other. These data also showed that the enthalpy and entropy changes of grouper antibody-hapten reactions resemble those reported for rabbit IgG antibodies to this hapten. It is thus suggested that, although considerable evolution of immunoglobulin classes has occurred between fish and rabbits, the antibody combining site may have remained relatively unchanged during a large part of evolutionary time.

Full Text

The Full Text of this article is available as a PDF (842.5 KB).

Selected References

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

  1. Ashman R. F., Metzger H. A Waldenström macroglobulin which binds nitrophenyl ligands. J Biol Chem. 1969 Jun 25;244(12):3405–3414. [PubMed] [Google Scholar]
  2. Clem I. W., De Boutaud F., Sigel M. M. Phylogeny of immunoglobulin structure and function. II. Immunoglobulins of the nurse shark. J Immunol. 1967 Dec;99(6):1226–1235. [PubMed] [Google Scholar]
  3. Clem L. W., Small P. A., Jr Phylogeny of immunoglobulin structure and function. I. Immunoglobulins of the lemon shark. J Exp Med. 1967 May 1;125(5):893–920. doi: 10.1084/jem.125.5.893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. EISEN H. N. EQUILIBRIUM DIALYSIS FOR MEASUREMENT OF ANTIBODY-HAPTEN AFFINITIES. Methods Med Res. 1964;10:106–114. [PubMed] [Google Scholar]
  5. EISEN H. N., SISKIND G. W. VARIATIONS IN AFFINITIES OF ANTIBODIES DURING THE IMMUNE RESPONSE. Biochemistry. 1964 Jul;3:996–1008. doi: 10.1021/bi00895a027. [DOI] [PubMed] [Google Scholar]
  6. FARAH F. S., KERN M., EISEN H. N. The preparation and some properties of purified antibody specific for the 2,4-dinitrophenyl group. J Exp Med. 1960 Dec 1;112:1195–1210. doi: 10.1084/jem.112.6.1195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hill W. C., Cebra J. J. Horse anti-SI immunoglobulins. I. Properties of gamma-M-antibody. Biochemistry. 1965 Dec;4(12):2575–2584. doi: 10.1021/bi00888a005. [DOI] [PubMed] [Google Scholar]
  8. Hodgins H. O., Weiser R. S., Ridgway G. J. The nature of antibodies and the immune response in rainbow trout (Salmo gairdneri). J Immunol. 1967 Sep;99(3):534–544. [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Leslie G. A., Clem L. W. Production of anti-hapten antibodies by several classes of lower vertebrates. J Immunol. 1969 Sep;103(3):613–617. [PubMed] [Google Scholar]
  11. Lindqvist K., Bauer D. C. Precipitin activity of rabbit macroglobulin antibody. Immunochemistry. 1966 Sep;3(5):373–384. doi: 10.1016/0019-2791(66)90175-3. [DOI] [PubMed] [Google Scholar]
  12. Lykakis J. J., Cox F. E. Immunological responses of the toad, Xenopus laevis, to the antigens of the ciliate, Tetrahymena pyriformis. Immunology. 1968 Sep;15(3):429–437. [PMC free article] [PubMed] [Google Scholar]
  13. Marchalonis J., Edelman G. M. Phylogenetic origins of antibody structure. II. Immunoglobulins in the primary immune response of the bullfrog, Rana catesbiana. J Exp Med. 1966 Nov 1;124(5):901–913. doi: 10.1084/jem.124.5.901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  15. Merler E., Karlin L., Matsumoto S. The valency of human gamma-M immunoglobulin antibody. J Biol Chem. 1968 Jan 25;243(2):386–390. [PubMed] [Google Scholar]
  16. Onoue K., Grossberg A. L., Yagi Y., Pressman D. Immunoglobulin M antibodies with ten combining sites. Science. 1968 Nov 1;162(3853):574–576. doi: 10.1126/science.162.3853.574. [DOI] [PubMed] [Google Scholar]
  17. Onoue K., Yagi Y., Grossberg A. L., Pressman D. Number of binding sites of rabbit macroglobulin antibody and its subunits. Immunochemistry. 1965 Dec;2(4):401–415. doi: 10.1016/0019-2791(65)90039-x. [DOI] [PubMed] [Google Scholar]
  18. Oudin J., Michel M. Idiotypy of rabbit antibodies. II. Comparison of idiotypy of various kinds of antibodies formed in the same rabbits against Salmonella typhi. J Exp Med. 1969 Sep 1;130(3):619–642. doi: 10.1084/jem.130.3.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Porath J., Axen R., Ernback S. Chemical coupling of proteins to agarose. Nature. 1967 Sep 30;215(5109):1491–1492. doi: 10.1038/2151491a0. [DOI] [PubMed] [Google Scholar]
  20. Sigel M. M., Clem L. W. Antibody response of fish to viral antigens. Ann N Y Acad Sci. 1965 Aug 10;126(1):662–677. doi: 10.1111/j.1749-6632.1965.tb14312.x. [DOI] [PubMed] [Google Scholar]
  21. Suran A. A., Tarail M. H., Papermaster B. W. Immunoglobulins of the leopard shark. I. Isolation and characterization of 17 S and 7 S immunoglobulins with precipitating activity. J Immunol. 1967 Oct;99(4):679–686. [PubMed] [Google Scholar]
  22. UHR J. W., FINKELSTEIN S., FRANKLIN E. C. Antibody response to bacteriophage phi-X-174 in non-mammalian vertebrates. Proc Soc Exp Biol Med. 1962 Oct;111:13–15. doi: 10.3181/00379727-111-27691. [DOI] [PubMed] [Google Scholar]
  23. Voss E. W., Jr, Russell W. J., Sigel M. M. Purification and binding properties of nurse shark antibody. Biochemistry. 1969 Dec;8(12):4866–4872. doi: 10.1021/bi00840a033. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES