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. 1976 Jan;73(1):118–122. doi: 10.1073/pnas.73.1.118

Subunit structure, cell surface orientation, and partial amino-acid sequences of murine histocompatibility antigens.

R Henning, R J Milner, K Reske, B A Cunningham, G M Edelman
PMCID: PMC335851  PMID: 54917

Abstract

Detergent and papain solubilized murine histocompatibility (H-2) antigens have been compared by gel exclusion chromatography, ultracentrifugation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and amino-acid sequence analysis. From these data, we propose a molecular model for the H-2 antigens that includes the size and arrangement of the subunits on the cell surface and in solution, and we provide evidence for the orientation of these molecules on the cell surface. Detergent solubilized H-2 antigens (molecular weight 116,000) consist of two disulfide-linked heavy chains (46,000 daltons) and two monocovalently associated light chains (12,000 daltons). Alkylation with iodoacetamide prior to extraction prevented the formation of a disulfide linkage between the two heavy chains. A water-soluble 51,000-dalton molecule (Fs) consisting of a 39,000-dalton fragment (FH) of the heavy chain and one intact light chain was obtained by papain digestion of cells or detergent extracts. Therefore, the disulfide linkage between the heavy chains is located in the remaining membrane-associated portion (Fm). Amino-acid sequence analysis of the FH fragment of H-2Kb by radiochemical techniques showed that it is identical to the detergent solubilized H-2Kb heavy chain in eight positions for the three amino acids tested. These data indicate that the fragment FH derives from the amino-terminus of the heavy chain and suggest that it projects outward from the cell surface, while the carboxyl-terminal region is associated with the plasma membrane. The described amino-terminal sequence data have been found constant in H-2Kb, H-2Kd, H-2Kk, H-2Db, and H-2Dd gene products. These data support the hypothesis that the K and D products of the major histocompatibility antigen complex have evolved by gene duplication.

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Selected References

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  1. Ackers G. K. Analytical gel chromatography of proteins. Adv Protein Chem. 1970;24:343–446. doi: 10.1016/s0065-3233(08)60245-4. [DOI] [PubMed] [Google Scholar]
  2. Bretscher M. S. Major human erythrocyte glycoprotein spans the cell membrane. Nat New Biol. 1971 Jun 23;231(25):229–232. doi: 10.1038/newbio231229a0. [DOI] [PubMed] [Google Scholar]
  3. Brown J. L., Kato K., Silver J., Nathenson S. G. Notable diversity in peptide composition of murine H-2K and H-2D alloantigens. Biochemistry. 1974 Jul 16;13(15):3174–3178. doi: 10.1021/bi00712a027. [DOI] [PubMed] [Google Scholar]
  4. Burnet F. M. A certain symmetry: histocompatibility antigens compared with immunocyte receptors. Nature. 1970 Apr 11;226(5241):123–126. doi: 10.1038/226123a0. [DOI] [PubMed] [Google Scholar]
  5. Cresswell P., Dawson J. R. Dimeric and monomeric forms of HL-A antigens solubilized by detergent. J Immunol. 1975 Jan;114(1 Pt 2):523–525. [PubMed] [Google Scholar]
  6. Cunningham B. A., Wang J. L., Berggård I., Peterson P. A. The complete amino acid sequence of beta 2-microglobulin. Biochemistry. 1973 Nov 20;12(24):4811–4822. doi: 10.1021/bi00748a001. [DOI] [PubMed] [Google Scholar]
  7. Fu S. M., Kunkel H. G. Membrane immunoglobulins of B lymphocytes: inability to detect certain characteristic IgM and IgD antigens. J Exp Med. 1974 Oct 1;140(4):895–903. doi: 10.1084/jem.140.4.895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gally J. A., Edelman G. M. The genetic control of immunoglobulin synthesis. Annu Rev Genet. 1972;6:1–46. doi: 10.1146/annurev.ge.06.120172.000245. [DOI] [PubMed] [Google Scholar]
  9. Grey H. M., Kubo R. T., Colon S. M., Poulik M. D., Cresswell P., Springer T., Turner M., Strominger J. L. The small subunit of HL-A antigens is beta 2-microglobulin. J Exp Med. 1973 Dec 1;138(6):1608–1612. doi: 10.1084/jem.138.6.1608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gunther G. R., Wang J. L., Yahara I., Cunningham B. A., Edelman G. M. Concanavalin A derivatives with altered biological activities. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1012–1016. doi: 10.1073/pnas.70.4.1012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Jacobs J. W., Kemper B., Niall H. D., Habener J. F., Potts J. T., Jr Structural analysis of human proparathyroid hormone by a new microsequencing approach. Nature. 1974 May 10;249(453):155–157. doi: 10.1038/249155a0. [DOI] [PubMed] [Google Scholar]
  13. Klein J., Shreffler D. C. The H-2 model for the major histocompatibility systems. Transplant Rev. 1971;6:3–29. doi: 10.1111/j.1600-065x.1971.tb00457.x. [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. Marchalonis J. J., Cone R. E., Santer V. Enzymic iodination. A probe for accessible surface proteins of normal and neoplastic lymphocytes. Biochem J. 1971 Oct;124(5):921–927. doi: 10.1042/bj1240921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McEwen C. R. Tables for estimating sedimentation through linear concentration gradients of sucrose solution. Anal Biochem. 1967 Jul;20(1):114–149. doi: 10.1016/0003-2697(67)90271-0. [DOI] [PubMed] [Google Scholar]
  17. Nakamuro K., Tanigaki N., Pressman D. Multiple common properties of human beta2-microglobulin and the common portion fragment derived from HL-A antigen molecules. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2863–2865. doi: 10.1073/pnas.70.10.2863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Peterson P. A., Cunningham B. A., Berggård I., Edelman G. M. 2 -Microglobulin--a free immunoglobulin domain. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1697–1701. doi: 10.1073/pnas.69.7.1697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Peterson P. A., Rask L., Sege K., Klareskog L., Anundi H., Ostberg L. Evolutionary relationship between immunoglobulins and transplantation antigens. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1612–1616. doi: 10.1073/pnas.72.4.1612. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. SANDERSON A. R. CYTOTOXIC REACTIONS OF MOUSE ISO-ANTISERA: PRELIMINARY CONSIDERATIONS. Br J Exp Pathol. 1964 Aug;45:398–408. [PMC free article] [PubMed] [Google Scholar]
  21. Schrader J. W., Cunningham B. A., Edelman G. M. Functional interactions of viral and histocompatibility antigens at tumor cell surfaces. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5066–5070. doi: 10.1073/pnas.72.12.5066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schwartz B. D., Kato K., Cullen S. E., Nathenson S. G. H-2 histocompatibility alloantigens. Some biochemical properties of the molecules solubilized by NP-40 detergent. Biochemistry. 1973 May 22;12(11):2157–2164. doi: 10.1021/bi00735a023. [DOI] [PubMed] [Google Scholar]
  23. Segrest J. P., Kahane I., Jackson R. L., Marchesi V. T. Major glycoprotein of the human erythrocyte membrane: evidence for an amphipathic molecular structure. Arch Biochem Biophys. 1973 Mar;155(1):167–183. doi: 10.1016/s0003-9861(73)80019-0. [DOI] [PubMed] [Google Scholar]
  24. Shearer G. M., Rehn T. G., Garbarino C. A. Cell-mediated lympholysis of trinitrophenyl-modified autologous lymphocytes. Effector cell specificity to modified cell surface components controlled by H-2K and H-2D serological regions of the murine major histocompatibility complex. J Exp Med. 1975 Jun 1;141(6):1348–1364. doi: 10.1084/jem.141.6.1348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Silver J., Hood L. Detergent-solubilised H-2 alloantigen is associated with a small molecular weight polypeptide. Nature. 1974 Jun 21;249(459):764–765. doi: 10.1038/249764a0. [DOI] [PubMed] [Google Scholar]
  26. Steck T. L. Cross-linking the major proteins of the isolated erythrocyte membrane. J Mol Biol. 1972 May 14;66(2):295–305. doi: 10.1016/0022-2836(72)90481-0. [DOI] [PubMed] [Google Scholar]
  27. Strominger J. L., Cresswell P., Grey H., Humphreys R. E., Mann D., McCuneJ, Parham P., Robb R., Sanderson A. R., Springer T. A. The immunoglobulin-like structure of human histocompatibility antigens. Transplant Rev. 1974;21(0):126–143. doi: 10.1111/j.1600-065x.1974.tb01549.x. [DOI] [PubMed] [Google Scholar]
  28. Summers M. R., Smythers G. W., Oroszlan S. Thin-layer chromatography of sub-nanomole amounts of phenylthiohydantoin (PTH) amino acids on polyamide sheets. Anal Biochem. 1973 Jun;53(2):624–628. doi: 10.1016/0003-2697(73)90114-0. [DOI] [PubMed] [Google Scholar]
  29. Svasti J., Milstein C. The complete amino acid sequence of a mouse kappa light chain. Biochem J. 1972 Jun;128(2):427–444. doi: 10.1042/bj1280427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tanford C., Nozaki Y., Reynolds J. A., Makino S. Molecular characterization of proteins in detergent solutions. Biochemistry. 1974 May 21;13(11):2369–2376. doi: 10.1021/bi00708a021. [DOI] [PubMed] [Google Scholar]
  31. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  32. Weiner A. M., Platt T., Weber K. Amino-terminal sequence analysis of proteins purified on a nanomole scale by gel electrophoresis. J Biol Chem. 1972 May 25;247(10):3242–3251. [PubMed] [Google Scholar]
  33. Zinkernagel R. M., Doherty P. C. Immunological surveillance against altered self components by sensitised T lymphocytes in lymphocytic choriomeningitis. Nature. 1974 Oct 11;251(5475):547–548. doi: 10.1038/251547a0. [DOI] [PubMed] [Google Scholar]

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