Abstract
Mouse lymphocyte H-2 and Ia glycoproteins have been analyzed with a two- dimensional (2-D) acrylamide gel electrophoresis technique, in which proteins are separated first according to their charge in isoelectrofocusing gels and then according to their size in sodium dodecyl sulfate gels. Individual polypeptide chains from radiolabeled cells are resolved as discrete spots on autoradiograms of the gels, forming patterns which are characteristic of the proteins in the sample. 2-D gels of H-2K, H-2D, and Ia glycoproteins immunoprecipitated from 35S-methionine-labeled cells reveal that these proteins exist in the cells as complex arrays of molecules heterogeneous in both size and charge. Lactoperoxidase-catalyzed radioiodination of lymphocyte surfaces labels only subsets of the total H-2 and Ia molecules with 125I, indicating that some of the molecules may represent cytoplasmic precursors of the cell surface proteins. This theory is supported by the kinetics of labeling of various spots in 35S-methionine pulse-chase experiments. The 2-D gel patterns obtained for both H-2 and Ia antigens have also been shown to be haplotype-specific and independent of the genetic background.
Full Text
The Full Text of this article is available as a PDF (3.0 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Barnstable C. J., Jones E. A., Bodmer W. F., Bodmer J. G., Arce-Gomez B., Snary D., Crumpton M. J. Genetics and serology of HL-A-linked human Ia antigens. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 2):443–455. doi: 10.1101/sqb.1977.041.01.052. [DOI] [PubMed] [Google Scholar]
- Cullen S. E., David C. S., Shreffler D. C., Nathenson S. G. Membrane molecules determined by the H-2 associated immune response region: isolation and some properties. Proc Natl Acad Sci U S A. 1974 Mar;71(3):648–652. doi: 10.1073/pnas.71.3.648. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cullen S. E., Nathenson S. G. Distribution of H-2 alloantigenic specificities on radiolabeled papain-solubilized antigen fragments. J Immunol. 1971 Aug;107(2):563–570. [PubMed] [Google Scholar]
- Cullen S. E., Schwartz B. D. An improved method for isolation of H-2 and Ia alloantigens with immunoprecipitation induced by protein A-bearing staphylococci. J Immunol. 1976 Jul;117(1):136–142. [PubMed] [Google Scholar]
- David C. S., Shreffler D. C., Frelinger J. A. New lymphocyte antigen system (Lna) controlled by the Ir region of the mouse H-2 complex. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2509–2514. doi: 10.1073/pnas.70.9.2509. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Démant P. H-2 gene complex and its role in alloimmune reactions. Transplant Rev. 1973;15:162–200. [PubMed] [Google Scholar]
- Frelinger J. A., Murphy D. B., McCormick J. F. Tla types of H-2 congenic and recombinant mice. Transplantation. 1974 Sep;18(3):292–294. [PubMed] [Google Scholar]
- Goding J. W., White E., Marchalonis J. J. Partial characterisation of Ia antigens on murine thymocytes. Nature. 1975 Sep 18;257(5523):230–231. doi: 10.1038/257230a0. [DOI] [PubMed] [Google Scholar]
- Götze D., Reisfeld R. A., Klein J. Serologic evidence for antigens controlled by the Ir region in mice. J Exp Med. 1973 Oct 1;138(4):1003–1008. doi: 10.1084/jem.138.4.1003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hansen T. H., Cullen S. E., Sachs D. H. Immunochemical evidence for an additional H-2 region closely linked to H-2D. J Exp Med. 1977 Feb 1;145(2):438–442. doi: 10.1084/jem.145.2.438. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hauptfeld V., Klein D., Klein J. Serological identification of an Ir-region product. Science. 1973 Jul 13;181(4095):167–169. doi: 10.1126/science.181.4095.167. [DOI] [PubMed] [Google Scholar]
- Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
- 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]
- McDevitt H. O., Benacerraf B. Genetic control of specific immune responses. Adv Immunol. 1969;11:31–74. doi: 10.1016/s0065-2776(08)60477-0. [DOI] [PubMed] [Google Scholar]
- McDevitt H. O., Deak B. D., Shreffler D. C., Klein J., Stimpfling J. H., Snell G. D. Genetic control of the immune response. Mapping of the Ir-1 locus. J Exp Med. 1972 Jun 1;135(6):1259–1278. doi: 10.1084/jem.135.6.1259. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McDevitt H. O., Delovitch T. L., Press J. L., Murphy D. B. Genetic and functional analysis of the Ia antigens: their possible role in regulating the immune response. Transplant Rev. 1976;30:197–235. doi: 10.1111/j.1600-065x.1976.tb00221.x. [DOI] [PubMed] [Google Scholar]
- Meo T., Krasteff T., Shreffler D. C. Immunochemical characterization of murine H-2 controlled Ss (serum substance) protein through identification of its human homologue as the fourth component of complement. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4536–4540. doi: 10.1073/pnas.72.11.4536. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Michaelson J., Flaherty L., Vitetta E., Poulik M. D. Molecular similarities between the Qa-2 alloantigen and other gene products of the 17th chromosome of the mouse. J Exp Med. 1977 Apr 1;145(4):1066–1070. doi: 10.1084/jem.145.4.1066. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murphy D. B., Herzenberg L. A., Okumura K., Herzenberg L. A., McDevitt H. O. A new I subregion (I-J) marked by a locus (Ia-4) controlling surface determinants on suppressor T lymphocytes. J Exp Med. 1976 Sep 1;144(3):699–712. doi: 10.1084/jem.144.3.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nathenson S. G., Cullen S. E. Biochemical properties and immunochemical-genetic relationships of mouse H-2 alloantigens. Biochim Biophys Acta. 1974 Apr 8;344(1):1–25. doi: 10.1016/0304-4157(74)90006-9. [DOI] [PubMed] [Google Scholar]
- Sachs D. H., Cone J. L. A mouse B-cell alloantigen determined by gene(s) linked to the major histocompatibility complex. J Exp Med. 1973 Dec 1;138(6):1289–1304. doi: 10.1084/jem.138.6.1289. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwartz B. D., Paul W. E., Shevach E. M. Guinea-pig Ia antigens: functional significance and chemical characterization. Transplant Rev. 1976;30:174–196. doi: 10.1111/j.1600-065x.1976.tb00220.x. [DOI] [PubMed] [Google Scholar]
- Shreffler D. C., David C. S., Cullen S. E., Frelinger J. A., Niederhuber J. E. Serological and functional evidence for further subdivision of the I regions of the H-2 gene complex. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 2):477–487. doi: 10.1101/sqb.1977.041.01.055. [DOI] [PubMed] [Google Scholar]
- Shreffler D. C., David C. S. The H-2 major histocompatibility complex and the I immune response region: genetic variation, function, and organization. Adv Immunol. 1975;20:125–195. doi: 10.1016/s0065-2776(08)60208-4. [DOI] [PubMed] [Google Scholar]
- Silver J., Cecka J. M., McMillan M., Hood L. Chemical characterization of products of the H-2 complex. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 1):369–377. doi: 10.1101/sqb.1977.041.01.044. [DOI] [PubMed] [Google Scholar]
- Silver J., Hood L. Structure and evolution of transplantation antigens: partial amino-acid sequences of H-2K and H-2D alloantigens. Proc Natl Acad Sci U S A. 1976 Feb;73(2):599–603. doi: 10.1073/pnas.73.2.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vitetta E. S., Capra J. D., Klapper D. G., Klein J., Uhr J. W. The partial amino-acid sequence of an H-2K molecule. Proc Natl Acad Sci U S A. 1976 Mar;73(3):905–909. doi: 10.1073/pnas.73.3.905. [DOI] [PMC free article] [PubMed] [Google Scholar]