Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1982 Apr 1;203(1):201–208. doi: 10.1042/bj2030201

Structurally inherent antigenic sites. Localization of the antigenic sites of the alpha-chain of human haemoglobin in three host species by a comprehensive synthetic approach.

A L Kazim, M Z Atassi
PMCID: PMC1158211  PMID: 6179509

Abstract

The antigenic structure of the alpha-chain of human haemoglobin was studied by a synthetic approach consisting of the synthesis of a series of consecutive overlapping peptides that together systematically represent the entire primary structure of the protein. This approach enabled the identification of a full profile of immunochemically active alpha-chain peptides and the localization of its major 'continuous' antigenic sites. Antibodies to haemoglobin raised in each of three different species (goat, rabbit and mouse) recognize similar sites on the alpha-chain. Further, the molecular locations of these sites coincide with alpha-chain regions extrapolated from antigenic sites of the conformationally similar myoglobin molecule. These findings support our earlier proposed concept of 'structurally inherent antigenic sites', namely that antigenicity is conferred on certain surface regions of proteins by virtue of their three-dimensional locations. Thus the antigenic sites of conformationally related proteins are likely to have similar molecular locations.

Full text

PDF
203

Selected References

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

  1. Atassi M. Z. Antigenic structure of myoglobin: the complete immunochemical anatomy of a protein and conclusions relating to antigenic structures of proteins. Immunochemistry. 1975 May;12(5):423–438. doi: 10.1016/0019-2791(75)90010-5. [DOI] [PubMed] [Google Scholar]
  2. Atassi M. Z. Chemical studies on haemoglobins A1 and A0. Biochem J. 1964 Oct;93(1):189–197. doi: 10.1042/bj0930189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Atassi M. Z., Kazim A. L. Distance calculation of residues neighbouring to lysozyme antigenic sites. Site-neighbouring residues whose evolutionary substitution can modify the characteristics and binding energy of the sites. Biochem J. 1980 Apr 1;187(1):163–172. doi: 10.1042/bj1870163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Atassi M. Z. Periodate oxidation of sperm-whale myoglobin and the role of the methionine residues in the antigen-antibody reaction. Biochem J. 1967 Feb;102(2):478–487. doi: 10.1042/bj1020478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Atassi M. Z. Precise determination of protein antigenic structures has unravelled the molecular immune recognition of proteins and provided a prototype for synthetic mimicking of other protein binding sites. Mol Cell Biochem. 1980 Aug 29;32(1):21–43. doi: 10.1007/BF00421293. [DOI] [PubMed] [Google Scholar]
  6. Atassi M. Z. Precise determination of the entire antigenic structure of lysozyme: molecular features of protein antigenic structures and potential of "surface-simulation" synthesis--a powerful new concept for protein binding sites. Immunochemistry. 1978 Dec;15(12):909–936. doi: 10.1016/0161-5890(78)90126-8. [DOI] [PubMed] [Google Scholar]
  7. Atassi M. Z., Sakata S., Kazim A. L. Localization and verification by synthesis of five antigenic sites of bovine serum albumin. Biochem J. 1979 May 1;179(2):327–331. doi: 10.1042/bj1790327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. BRAUNITZER G., GEHRING-MUELLER R., HILSCHMANN N., HILSE K., HOBOM G., RUDLOFF V., WITTMANN-LIEBOLD B. [The structure of normal adult human hemoglobins]. Hoppe Seylers Z Physiol Chem. 1961 Sep 20;325:283–286. doi: 10.1515/bchm2.1961.325.1.283. [DOI] [PubMed] [Google Scholar]
  10. Crowley J. B., Birkmeyer R. C., Tan-Wilson A. L. Additional antigenic sites on human hemoglobin evident upon separation of antibody populations. Mol Immunol. 1980 Dec;17(12):1449–1457. doi: 10.1016/0161-5890(80)90170-4. [DOI] [PubMed] [Google Scholar]
  11. Dean J., Schechter A. N. Conformation-specific antibodies to the alpha chain COOH terminus of hemoglobin A0. J Biol Chem. 1979 Sep 25;254(18):9185–9193. [PubMed] [Google Scholar]
  12. Fermi G. Three-dimensional fourier synthesis of human deoxyhaemoglobin at 2-5 A resolution: refinement of the atomic model. J Mol Biol. 1975 Sep 15;97(2):237–256. doi: 10.1016/s0022-2836(75)80037-4. [DOI] [PubMed] [Google Scholar]
  13. Geraci G., Parkhurst L. J., Gibson Q. H. Preparation and properties of alpha- and beta-chains from human hemoglobin. J Biol Chem. 1969 Sep 10;244(17):4664–4667. [PubMed] [Google Scholar]
  14. Gray G. D., Mickelson M. M., Crim J. A. The demonstration of two gamma-globulin subclasses in the goat. Immunochemistry. 1969 Sep;6(5):641–644. doi: 10.1016/0019-2791(67)90128-0. [DOI] [PubMed] [Google Scholar]
  15. HILL R. J., KONIGSBERG W. The structure of human hemoglobin. IV. The chymotryptic digestion of the alpha chain of human hemoglobin. J Biol Chem. 1962 Oct;237:3151–3156. [PubMed] [Google Scholar]
  16. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  17. Hurrell J. G., Smith J. A., Leach S. J. The detection of five antigenically reactive regions in the soybean leghemoglobin a molecule. Immunochemistry. 1978 May;15(5):297–302. doi: 10.1016/0161-5890(78)90089-5. [DOI] [PubMed] [Google Scholar]
  18. Kazim A. L., Atassi M. Z. A novel and comprehensive synthetic approach for the elucidation of protein antigenic structures. Determination of the full antigenic profile of the alpha-chain of human haemoglobin. Biochem J. 1980 Oct 1;191(1):261–264. doi: 10.1042/bj1910261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kazim A. L., Atassi M. Z. Antibodies against protein antigenic sites that are identical in the homologous protein of the immunized animal. Autoreactivity in rabbits of antibodies to sperm-whale myoglobin. Biochim Biophys Acta. 1977 Sep 27;494(1):277–282. doi: 10.1016/0005-2795(77)90156-8. [DOI] [PubMed] [Google Scholar]
  20. Kazim A. L., Atassi M. Z. Haemoglobin binding with haptoglobin. Unequivocal demonstration that the beta-chains of human haemoglobin bind to haptoglobin. Biochem J. 1980 Jan 1;185(1):285–287. doi: 10.1042/bj1850285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kazim A. L., Atassi M. Z. Nearest-neighbour analysis of myoglobin antigenic sites. Nearest-neighbour residues whose replacement can alter the environment of binding-site residue(s) and thus change their characteristics and binding capability. Biochem J. 1980 Dec 1;191(3):673–680. doi: 10.1042/bj1910673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kazim A. L., Atassi M. Z. Prediction and conformation by synthesis of two antigenic sites in human haemoglobin by extrapolation from the known antigenic structure of sperm-whale myoglobin. Biochem J. 1977 Oct 1;167(1):275–278. doi: 10.1042/bj1670275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Koketsu J., Atassi M. Z. Immunochemistry of sperm-whale myoglobin. XVI. Accurate delineation of the single region in sequence 1-55 by immunochemical studies of synthetic peptides. Some conclusions concerning antigenic structures of proteins. Immunochemistry. 1974 Jan;11(1):1–8. doi: 10.1016/0019-2791(74)90335-8. [DOI] [PubMed] [Google Scholar]
  24. Krco C. J., Kazim A. L., Atassi M. Z., David C. S. Genetic control of the immune response to haemoglobin. II. Studies using purified alpha-chain and beta-chain as immunogens. J Immunogenet. 1981 Oct;8(5):395–403. doi: 10.1111/j.1744-313x.1981.tb00944.x. [DOI] [PubMed] [Google Scholar]
  25. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  26. Lee C. L., Pai R. C., Atassi M. Z. Enzymic and immunochemical properties of lysozyme--XV. Delineation of the reactive site around the two central disulfides by immunochemical studies of novel synthetic peptides that contain diglycyl bridges instead of disulfides. Immunochemistry. 1976 Aug;13(8):681–687. doi: 10.1016/0019-2791(76)90209-3. [DOI] [PubMed] [Google Scholar]
  27. Liu T. Y., Chang Y. H. Hydrolysis of proteins with p-toluenesulfonic acid. Determination of tryptophan. J Biol Chem. 1971 May 10;246(9):2842–2848. [PubMed] [Google Scholar]
  28. March S. C., Parikh I., Cuatrecasas P. A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal Biochem. 1974 Jul;60(1):149–152. doi: 10.1016/0003-2697(74)90139-0. [DOI] [PubMed] [Google Scholar]
  29. Matzku S., Zöller M. Iodination of immunoadsorbed antibodies: chloramine T vs lactoperoxidase. Immunochemistry. 1977 May;14(5):367–371. doi: 10.1016/0019-2791(77)90238-5. [DOI] [PubMed] [Google Scholar]
  30. Perutz M. F., Muirhead H., Cox J. M., Goaman L. C. Three-dimensional Fourier synthesis of horse oxyhaemoglobin at 2.8 A resolution: the atomic model. Nature. 1968 Jul 13;219(5150):131–139. doi: 10.1038/219131a0. [DOI] [PubMed] [Google Scholar]
  31. Reichlin M. Amino acid substitution and the antigenicity of globular proteins. Adv Immunol. 1975;20:71–123. doi: 10.1016/s0065-2776(08)60207-2. [DOI] [PubMed] [Google Scholar]
  32. Takagaki Y., Hirayama A., Fujio H., Amano T. Antibodies to a continuous region at residues 38-54 of hen egg white lysozyme found in a small fraction of anti-hen egg white lysozyme antibodies. Biochemistry. 1980 May 27;19(11):2498–2505. doi: 10.1021/bi00552a031. [DOI] [PubMed] [Google Scholar]
  33. Tan-Wilson A. L., Reichlin M., Noble R. W. Properties of goat anti-human hemoglobin antibodies fractionated on subunit affinity columns. Immunochemistry. 1976 Jun;13(6):491–498. doi: 10.1016/0019-2791(76)90324-4. [DOI] [PubMed] [Google Scholar]
  34. Twining S. S., Atassi M. Z. Use of immunoadsorbents for the study of antibody binding to sperm whale myoglobin and its synthetic antigenic sites. J Immunol Methods. 1979;30(2):139–151. doi: 10.1016/0022-1759(79)90088-7. [DOI] [PubMed] [Google Scholar]
  35. Twining S. S., Lehmann H., Atassi M. Z. The antibody response to myoglobin is independent of the immunized species. Analysis in terms of replacements in the antigenic sites and in environmental residues of the cross-reactions of fifteen myoglobins with sperm-whale myoglobin antisera raised in different species. Biochem J. 1980 Dec 1;191(3):681–697. doi: 10.1042/bj1910681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Visscher M. B. Letter: Faculty opinion on changes in medical education. JAMA. 1974 Jul 1;229(1):22–23. doi: 10.1001/jama.1974.03230390014004. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES