Abstract
Assembly of HLA class I molecules was studied using pulse-chase labeling of B-lymphoblastoid cell lines with 35S-methionine, immunoprecipitation with antibodies detecting free or β2-microglobulin-associated heavy chain and isoelectric focusing. Marked differences between the products of different class I alleles were noted. HLA-B51 assembled very inefficiently, with considerable free heavy chain still detected in an unsialated form after a four hour chase. The closely related molecule HLA-B35 was in contrast rapidly assembled, all newly synthesized heavy chain being detected in a β2m-associated, sialated form within 30 minutes. Analysis of naturally occurring variants related to HLA-B35 and HLA-B51 localized the region determining assembly efficency to the α2 domain, in which these molecules differ at eight amino acid residues. The effect was not due to a linked dominant gene, as both patterns of assembly were observed in a single cell line.
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Allen, H., Fraser, J., Flyer, S., Calvin, S., and Flavell, R.: β2-microglobulin is not required for cell surface expression of the murine class I histocompatibility antigen H-2Db or of a truncated H-2Db. Proc Natl Acad Sci USA 79: 7447, 1988
Allsopp, C. E. M., Hill, A. V. S., Kwiatowski, D., Hughes, A., Bunce, M., Taylor, C. J., Pazmany, L., Brewster, D., McMichael, A. J., and Greenwood, B. M.: Sequence analysis of HLA-Bw53, a common West African allele, suggests an origin by gene conversion of HLA-B35. Hum Immunol 30: 105–109, 1991
Barnstable, C. J., Bodmer, W., Brown, G. E. A., Galfre, G., Milstein, C., Williams, A. F., and Ziegler, A.: Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens — new tools for genetic analysis. Cell 14: 9, 1978
Beck, J. C., Hansen, T. H., Cullen, S. E., and Lee, D. R.: Slower processing, weaker β2m association, and lower surface expression of H-2Ld are influenced by its amino terminus. J Immunol 137: 916–923, 1986
Bjorkman, P. J. and Parham, P.: Structure, function, and diversity of class I major histocompatibility complex molecules. Annu Rev Biochem 59: 253–288, 1990
Bjorkman, P. J., Saper, M. A., Samraoui, B., Bennett, W. S., Strominger, J. L., and Wiley, D. C.: The foreign antigen binding site and T cell recognition regions of class I histocompatibility antigens. Nature 329: 512–518, 1987
Chen, B. P., Lam, V., Kraus, E. E., DeMars, R., and Sondel, P. M.: Restriction of Epstein-Barr Virus-specific cytotoxic T cells by HLA-A,-B, and-C molecules. Hum Immunol 26: 137–147, 1989
Cox, J. H., Bennink, J. R., and Yewdell, J. W.: Retention of Adenovirus E19 Glycoprotein in the endoplasmic reticulum is essential to its ability to block antigen presentation. J Exp Med 174: 1629–1637, 1991
Degen, E. and Williams, D. B.: Participation of a novel 88-kD protein in the biogenesis of murine class I histocompatibility molecules. J Cell Biol 112: 1099–1115, 1991
Doherty, P. C., Biddison, W. E., Bennink, J. R., and Knowles, B. B.: Cytotoxic T cell responses in mice infected with influenza and vaccinia viruses vary in magnitude with H-2 genotype. J exp Med 148: 534–543, 1978
Doherty, P. C. and Zinkernagel, R. M.: A biological role for the major histocompatibility antigens. Lancet i: 1406–1409, 1975
Elliot, T., Cerundolo, V., Elvin, J., and Townsend, A.: Peptide-induced conformational change of the class I heavy chain. Nature 351: 402–406, 1991
Emerson, S. G., Murphy, D. B., and Cone, R. E.: Selective turnover and shedding of H-2K and H-2D antigens is controlled by the major histocompatibility complex. J Exp Med 152: 783–795, 1980
Falk, K., Rotzschke, O., Stevanovic, S., Jung, G., and Rammensee, H.-G.: Allele specific motifs revealed by sequencing of self peptides elluted from MHC molecules. Nature 351: 290–296, 1991
Gething, M. J. and Sambrook, J.: Protein folding in the cell. Nature 355: 33–45, 1992
Gussow, D., Rein, R. S., Meijer, I., de Hoog, W., Seemann, G. H. A., Hochstenbach, F. M., and Ploegh, H. L.: Isolation, expression, and the primary structure of HLA-Cw1 and HLA-Cw2 genes: evolutionary aspects. Immunogenetics 25: 213–322, 1987
Hansen, T. H., Myers, N. B., and Lee, D. R.: Studies of two antigenic forms of Ld with disparate β2-microglobulin (β2m) associations suggest that β2m facilitates the folding of the α1 and α2 domains during de novo synthesis. J Immunol 140: 3522–3529, 1988
Hayashi, H., Ennis, P. D., Ariga, H., Salter, R. D., Parham, P., Kano, K., and Takiguchi, M.: HLA-B51 and HLA-Bw52 differ by only two amino acids which are in the helical region of the alpha 1 domain. J Immunol 142: 306–311, 1989
Hayashi, H., Ooba, T., Nakayama, S., Sekimata, M., Kano, K., and Takiguchi, M.: Allospecificities between HLA-Bw53 and HLA-B35 are generated by substitution of the residues associated with HLA-Bw4/Bw6 public epitopes. Immunogenetics 32: 195–199, 1990
Jeffries, W. A. and Burgert, H.-G.: E3/19K from Adenovirus 2 is an immunosubsersive protein that binds to a structural motif regulating the intracellular transport of major histocompatibility complex class I proteins. J Exp Med 172: 1653–1664, 1990
Klar, D. and Hammerling, G. J.: Induction of assembly of MHC class I heavy chains with β2 microglobulin by interferon-γ. EMBO J 8: 475–481, 1989
Lawlor, D. A., Zemmour, J., Ennis, P. D., and Parham, P.: Evolution of Class-I MHC genes and proteins: from natural selection to thymic selection. Annu Rev Immunol 8: 23–63, 1990
Lie, W.-R., Myers, N. B., Gorka, J., Rubocki, R. J., Connolly, J. M., and Hansen, T. H.: Peptide ligand-induced conformation and surface expression of the Ld class I MHC molecule. Nature 344: 439–441, 1990
Livingstone, A. M., Powis, S. J., Diamond, A. G., Butcher, G. W., and Howard, J. C.: A trans-acting major histocompatibility complex-linked gene whose alleles determine gain and loss changes in the antigenic structure of a classical class I molecule. J Exp Med 170: 777–795, 1989
Madden, D. R., Gorga, J. C., Strominger, J. L., and Wiley, D. C.: The structure of HLA-B27 reveals nonamer “self-peptides” bound in an extended conformation. Nature 353: 321–325, 1991
McMichael, A., Gotch, F., and Rothbard, J.: HLA-B37 determines an influenza A virus nucleoprotein epitope recognized by cytotoxic T lymphocytes. J Exp Med 164: 1397–1406, 1986
Myers, N. B., Lie, W.-R., Nett, M., Rubocki, R. J., and Hansen, T. H.: The conformation of Ld induced by β2-microglobulin is fixed during de novo synthesis and are reversible by exchange or dissociation. J Immunol 142: 2751–2758, 1989
Neefjes, J. J., Breur-Vriesendorp, B. S., van-Seventer, G. A., Ivanyi, P., and Ploegh, H. L.: An improved biochemical method for the analysis of HLA-Class I antigens. Definition of new HLA-Class I subtypes. Hum Immunol 16: 169–181, 1986
Neefjes, J. J. and Ploegh, H. L.: Allele and locus-specific differences in cell surface expression and the association of HLA class I heavy chain with β2-microglobulin: differential effects of inhibition of glycosylation on class I subunit association. Eur J Immunol 18: 801–810, 1988
Nuchtern, J. G., Bonifacino, J. S., Biddison, W. E., and Klausner, R. D.: Brefeldin A implicates egress from the endoplasmic reticulum in class I-restricted antigen presentation. Nature 339: 223–226, 1989
Ooba, T., Hayashi, H., Karaki, S., Tanabe, M., Kano, K., and Takiguchi, M.: The structure of HLA-B35 suggests that it is derived from HLA-Bw58 by two genetic mechanisms. Immunogenetics 30: 76–80, 1989
Pala, P. and Askonas, B. A.: Low responder MHC alleles for Tc recognition of influenza nucleoprotein. Immunogenetics 23: 379–384, 1986
Powis, S. J., Howard, J. C., and Butcher, G. W.: The major histocompatibility complex class II-linked cim locus controls the kinetics of intracellular transport of a classical class I molecule. J Exp Med 173: 913–921, 1991
Sekimata, M., Hiraiwa, M., Andrien, M., Dupont, E., Karaki, S., Yamamoto, J., Kano, K., and Takiguchi, M.: Allodeterminants and evolution of a novel HLA-B5 CREG antigen, HLA-B SNA. J Immunol 144: 3228–3233, 1990
Scheisl, B.: HLA-Typing: Problems and Solutions. Biotest Diagnostics (Biotest-Mitteilungen, Biotest, Dreieich, FRG), 1987
Snary, D., Barnstable, C. J., Bodmer, W. F., and Crumpton, M. J.: Molecular structure of human histocompatibility antigens: the HLA-C series. Eur J Immunol 7: 580–585, 1977
Stam, N., Vroom, T., Peters, P., nPastoors, E., and Ploegh, H.: HLA-A and HLA-B specific monoclonal antibodies reactive with free heavy chains in Western blots, in formalin-fixed paraffin-embedded tissue sections and in cryo-immuno-electron microscopy. Int Immunol 2: 113–125, 1990
Stam, N. J., Spits, H., and Ploegh, H. L.: Monoclonal antibodies raised against denatured HLA-B locus heavy chains permit biochemical characterization of certain HLA-C locus products. J Immunol 137: 2299, 1986
Townsend, A., Elliott, T., Cerundolo, V., Foster, L., Barber, B., and Tse, A.: Assembly of MHC class I molecules analyzed in vitro. Cell 62: 285–295, 1990
Townsend, A., Ohlen, C., Bastin, J., Ljunggren, H. G., Foster, L., and Karre, K.: Association of class I major histocompatibility heavy and light chains induced by viral peptides. Nature 340: 443–448, 1989
Weis, J. H. and Murre, C.: Differential Expression of H-2Dd and H-2Ld Histocompatibility Antigens. J Exp Med 161: 356–365, 1985
Williams, D. B., Swiedler, S. J., and Hart, G. W.: Intracellular transport of membrane glycoproteins: two closely related histocompatibility antigens differ in their rates of transit to the cell surface. J Cell Biol 101: 725–734, 1985
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Hill, A., Takiguchi, M. & McMichael, A. Different rates of HLA class I molecule assembly which are determined by amino acid sequence in the α2 domain. Immunogenetics 37, 95–101 (1993). https://doi.org/10.1007/BF00216831
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DOI: https://doi.org/10.1007/BF00216831