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
. 1979 Nov;76(11):5848–5852. doi: 10.1073/pnas.76.11.5848

Effect of interchain disulfide bond on hapten binding properties of light chain dimer of protein 315.

R Zidovetski, A Licht, I Pecht
PMCID: PMC411749  PMID: 118454

Abstract

The hapten binding characteristics of the covalent light chain dimer, derived from the murine IgA secreted by plasmacytoma MOPC-315, to two nitroaromatic compounds, epsilon-N-(2,4-dinitrophenyl)-L-lysine and 4-(alpha-N-alanine)-m-nitrobenz-2-oxa-1,3-diazole, were investigated by differential spectroscopic titrations. The binding curves for both haptens were found to display sigmoidity similar to that reported earlier for the reduced and alkylated dimer held together by noncovalent bonds only. However, the presence of the interchain disulfide bond in the covalent dimer was found to cause marked changes in its binding properties. The data, like those obtained for the noncovalent dimer, fit the allosteric model of Monod, Wyman, and Changeux in which binding of the first hapten to the dimer causes a conversion of both sites of the protein molecule from a lower to a higher affinity conformation. However, the binding parameters show that both the affinity and the positive cooperativity in the interaction between haptens and the covalent dimer are significantly enhanced. The differences in the parameters of the binding and of the allosteric transition caused by the presence of the interchain disulfide bond demonstrate the existence of longitudinal interactions in immunoglobulin derivatives. These properties of the light chain dimer make it a potential model for the receptors present on thymus-derived lymphocytes.

Full text

PDF
5849

Images in this article

5849Image
on p.5849

Selected References

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

  1. Abel C. A., Grey H. M. Studies on the structure of mouse gamma-A myeloma proteins. Biochemistry. 1968 Jul;7(7):2682–2688. doi: 10.1021/bi00847a035. [DOI] [PubMed] [Google Scholar]
  2. Blanden R. V., Ada G. L. A dual recognition model for cytotoxic T cells based on thymic selection of precursors with low affinity for Self H-2 antigens. Scand J Immunol. 1978 Mar;7(3):181–190. doi: 10.1111/j.1365-3083.1978.tb00442.x. [DOI] [PubMed] [Google Scholar]
  3. Edmundson A. B., Ely K. R., Girling R. L., Abola E. E., Schiffer M., Westholm F. A., Fausch M. D., Deutsch H. F. Binding of 2,4-dinitrophenyl compounds and other small molecules to a crystalline lambda-type Bence-Jones dimer. Biochemistry. 1974 Aug 27;13(18):3816–3827. doi: 10.1021/bi00715a031. [DOI] [PubMed] [Google Scholar]
  4. Eisen H. N., Simms E. S., Potter M. Mouse myeloma proteins with antihapten antibody acitivity. The protein produced by plasma cell tumor MOPC-315. Biochemistry. 1968 Nov;7(11):4126–4134. doi: 10.1021/bi00851a048. [DOI] [PubMed] [Google Scholar]
  5. Ely K. R., Firca J. R., Williams K. J., Abola E. E., Fenton J. M., Schiffer M., Panagiotopoulos N. C., Edmundson A. B. Crystal properties as indicators of conformational changes during ligand binding or interconversion of Mcg light chain isomers. Biochemistry. 1978 Jan 10;17(1):158–167. doi: 10.1021/bi00594a023. [DOI] [PubMed] [Google Scholar]
  6. Epp O., Lattman E. E., Schiffer M., Huber R., Palm W. The molecular structure of a dimer composed of the variable portions of the Bence-Jones protein REI refined at 2.0-A resolution. Biochemistry. 1975 Nov 4;14(22):4943–4952. doi: 10.1021/bi00693a025. [DOI] [PubMed] [Google Scholar]
  7. Firca J. R., Ely K. R., Kremser P., Westholm F. A., Dorrington K. J., Edmundson A. B. Interconversion of conformational isomers of light chains in the Mcg immunoglobulins. Biochemistry. 1978 Jan 10;17(1):148–158. doi: 10.1021/bi00594a022. [DOI] [PubMed] [Google Scholar]
  8. Goetzl E. J., Metzger H. Affinity labeling of a mouse myeloma protein which binds nitrophenyl ligands. Kinetics of labeling and isolation of a labeled peptide. Biochemistry. 1970 Mar 3;9(5):1267–1278. doi: 10.1021/bi00807a031. [DOI] [PubMed] [Google Scholar]
  9. Haber J. E., Koshland D. E., Jr Relation of protein subunit interactions to the molecular species observed during cooperative binding of ligands. Proc Natl Acad Sci U S A. 1967 Nov;58(5):2087–2093. doi: 10.1073/pnas.58.5.2087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Janeway C. A., Wigzell H., Binz H. Two different VH gene products make up the T-cell receptors. Scand J Immunol. 1976;5(9):993–1001. doi: 10.1111/j.1365-3083.1976.tb03051.x. [DOI] [PubMed] [Google Scholar]
  11. Klein M., Kells D. I., Tinker D. O., Dorrington K. J. Thermodynamic and conformational studies on an immunoglobulin light chain which reversibly precipitates at low temperatures. Biochemistry. 1977 Feb 8;16(3):552–560. doi: 10.1021/bi00622a032. [DOI] [PubMed] [Google Scholar]
  12. Klein M., Kortan C., Kells D. I., Dorrington K. J. Equilibrium and kinetic aspects of the interaction of isolated variable and constant domains of light chain with the Fd' fragment of immunoglobulin G. Biochemistry. 1979 Apr 17;18(8):1473–1481. doi: 10.1021/bi00575a014. [DOI] [PubMed] [Google Scholar]
  13. Koshland D. E., Jr, Némethy G., Filmer D. Comparison of experimental binding data and theoretical models in proteins containing subunits. Biochemistry. 1966 Jan;5(1):365–385. doi: 10.1021/bi00865a047. [DOI] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lancet D., Licht A., Schechter I., Pecht I. Hapten-induced allosteric transition in the light chain dimer of an immunoglobulin. Nature. 1977 Oct 27;269(5631):827–829. doi: 10.1038/269827a0. [DOI] [PubMed] [Google Scholar]
  16. Lancet D., Pecht I. Kinetic evidence for hapten-induced conformational transition in immunoglobin MOPC 460. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3549–3553. doi: 10.1073/pnas.73.10.3549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lancet D., Pecht I. Spectroscopic and immunochemical studies with nitrobenzoxadiazolealanine, a fluorescent dinitrophenyl analogue. Biochemistry. 1977 Nov 15;16(23):5150–5157. doi: 10.1021/bi00642a031. [DOI] [PubMed] [Google Scholar]
  18. Licht A., Lancet D., Schechter I., Pecht I. Thermodynamic and spectroscopic comparison of the binding sites of the mouse myeloma protein 315 and of its light chain dimer. FEBS Lett. 1977 Jun 15;78(2):211–215. doi: 10.1016/0014-5793(77)80308-6. [DOI] [PubMed] [Google Scholar]
  19. Little J. R., Eisen H. N. Evidence for tryptophan in the active sites of antibodies to polynitrobenzenes. Biochemistry. 1967 Oct;6(10):3119–3125. doi: 10.1021/bi00862a020. [DOI] [PubMed] [Google Scholar]
  20. MONOD J., WYMAN J., CHANGEUX J. P. ON THE NATURE OF ALLOSTERIC TRANSITIONS: A PLAUSIBLE MODEL. J Mol Biol. 1965 May;12:88–118. doi: 10.1016/s0022-2836(65)80285-6. [DOI] [PubMed] [Google Scholar]
  21. Padlan E. A., Davies D. R., Pecht I., Givol D., Wright C. Model-building studies of antigen-binding sites: the hapten-binding site of mopc-315. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 2):627–637. doi: 10.1101/sqb.1977.041.01.072. [DOI] [PubMed] [Google Scholar]
  22. Poljak R. J., Amzel L. M., Phizackerley R. P. Studies on the three-dimensional structure of immunoglobulins. Prog Biophys Mol Biol. 1976;31(1):67–93. doi: 10.1016/0079-6107(78)90005-6. [DOI] [PubMed] [Google Scholar]
  23. Schechter I., Ziv E. Binding of 2,4-dinitrophenyl derivatives by the light chain dimer obtained from immunoglobulin A produced by MOPC-315 mouse myeloma. Biochemistry. 1976 Jun 29;15(13):2785–2790. doi: 10.1021/bi00658a013. [DOI] [PubMed] [Google Scholar]
  24. Shearer G. M., Rehn T. G., Schmitt-Verhulst A. M. Role of the murine major histocompatibility complex in the specificity of in vitro T-cell-mediated lympholysis against chemically-modified autologous lymphocytes. Transplant Rev. 1976;29:222–246. doi: 10.1111/j.1600-065x.1976.tb00203.x. [DOI] [PubMed] [Google Scholar]
  25. Stevenson G. T. The binding of haptens by the polypeptide chains of rabbit antibody molecules. Biochem J. 1973 Aug;133(4):827–836. doi: 10.1042/bj1330827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Underdown B. J., Simms E. S., Eisen H. N. Subunit structure and number of combining sites of the immunoglobulin A myeloma protein produced by mouse plasmacytoma MOPC-315. Biochemistry. 1971 Nov 23;10(24):4359–4368. doi: 10.1021/bi00800a002. [DOI] [PubMed] [Google Scholar]
  27. Vuk-Pavlović S., Blatt Y., Glaudemans C. P., Lancet D., Pecht I. Hapten-linked conformational equilibria in immunolglobulins XRPC-24 and J-539 observed by chemical relaxation. Biophys J. 1978 Oct;24(1):161–174. doi: 10.1016/S0006-3495(78)85353-3. [DOI] [PMC free article] [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