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. 1996 Feb 1;183(2):527–534. doi: 10.1084/jem.183.2.527

An HLA-A2-restricted tyrosinase antigen on melanoma cells results from posttranslational modification and suggests a novel pathway for processing of membrane proteins

PMCID: PMC2192446  PMID: 8627164

Abstract

T lymphocytes recognize antigens consisting of peptides presented by class I and II major histocompatibility complex (MHC) molecules. The peptides identified so far have been predictable from the amino acid sequences of proteins. We have identified the natural peptide target of a CTL clone that recognizes the tyrosinase gene product on melanoma cells. The peptide results from posttranslational conversion of asparagine to aspartic acid. This change is of central importance for peptide recognition by melanoma-specific T cells, but has no impact on peptide binding to the MHC molecule. This posttranslational modification has not been previously described for any MHC-associated peptide and represents the first demonstration of posttranslational modification of a naturally processed class I-associated peptide. This observation is relevant to the identification and prediction of potential peptide antigens. The most likely mechanism for production of this peptide leads to the suggestion that antigenic peptides can be derived from proteins that are translated into the endoplasmic reticulum.

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Selected References

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  1. Aronson N. N., Jr, Kuranda M. J. Lysosomal degradation of Asn-linked glycoproteins. FASEB J. 1989 Dec;3(14):2615–2622. doi: 10.1096/fasebj.3.14.2531691. [DOI] [PubMed] [Google Scholar]
  2. Bakker A. B., Schreurs M. W., de Boer A. J., Kawakami Y., Rosenberg S. A., Adema G. J., Figdor C. G. Melanocyte lineage-specific antigen gp100 is recognized by melanoma-derived tumor-infiltrating lymphocytes. J Exp Med. 1994 Mar 1;179(3):1005–1009. doi: 10.1084/jem.179.3.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bednarek M. A., Sauma S. Y., Gammon M. C., Porter G., Tamhankar S., Williamson A. R., Zweerink H. J. The minimum peptide epitope from the influenza virus matrix protein. Extra and intracellular loading of HLA-A2. J Immunol. 1991 Dec 15;147(12):4047–4053. [PubMed] [Google Scholar]
  4. Bertoletti A., Chisari F. V., Penna A., Guilhot S., Galati L., Missale G., Fowler P., Schlicht H. J., Vitiello A., Chesnut R. C. Definition of a minimal optimal cytotoxic T-cell epitope within the hepatitis B virus nucleocapsid protein. J Virol. 1993 Apr;67(4):2376–2380. doi: 10.1128/jvi.67.4.2376-2380.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bodmer H., Ogg G., Gotch F., McMichael A. Anti-HLA-A2 antibody-enhancement of peptide association with HLA-A2 as detected by cytotoxic T lymphocytes. Nature. 1989 Nov 23;342(6248):443–446. doi: 10.1038/342443a0. [DOI] [PubMed] [Google Scholar]
  6. Boon T., Cerottini J. C., Van den Eynde B., van der Bruggen P., Van Pel A. Tumor antigens recognized by T lymphocytes. Annu Rev Immunol. 1994;12:337–365. doi: 10.1146/annurev.iy.12.040194.002005. [DOI] [PubMed] [Google Scholar]
  7. Bouchard B., Fuller B. B., Vijayasaradhi S., Houghton A. N. Induction of pigmentation in mouse fibroblasts by expression of human tyrosinase cDNA. J Exp Med. 1989 Jun 1;169(6):2029–2042. doi: 10.1084/jem.169.6.2029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brichard V., Van Pel A., Wölfel T., Wölfel C., De Plaen E., Lethé B., Coulie P., Boon T. The tyrosinase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med. 1993 Aug 1;178(2):489–495. doi: 10.1084/jem.178.2.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chen Y., Sidney J., Southwood S., Cox A. L., Sakaguchi K., Henderson R. A., Appella E., Hunt D. F., Sette A., Engelhard V. H. Naturally processed peptides longer than nine amino acid residues bind to the class I MHC molecule HLA-A2.1 with high affinity and in different conformations. J Immunol. 1994 Mar 15;152(6):2874–2881. [PubMed] [Google Scholar]
  10. Chicz R. M., Urban R. G., Gorga J. C., Vignali D. A., Lane W. S., Strominger J. L. Specificity and promiscuity among naturally processed peptides bound to HLA-DR alleles. J Exp Med. 1993 Jul 1;178(1):27–47. doi: 10.1084/jem.178.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Coulie P. G., Brichard V., Van Pel A., Wölfel T., Schneider J., Traversari C., Mattei S., De Plaen E., Lurquin C., Szikora J. P. A new gene coding for a differentiation antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med. 1994 Jul 1;180(1):35–42. doi: 10.1084/jem.180.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cox A. L., Skipper J., Chen Y., Henderson R. A., Darrow T. L., Shabanowitz J., Engelhard V. H., Hunt D. F., Slingluff C. L., Jr Identification of a peptide recognized by five melanoma-specific human cytotoxic T cell lines. Science. 1994 Apr 29;264(5159):716–719. doi: 10.1126/science.7513441. [DOI] [PubMed] [Google Scholar]
  13. Engelhard V. H., Appella E., Benjamin D. C., Bodnar W. M., Cox A. L., Chen Y., Henderson R. A., Huczko E. L., Michel H., Sakaguichi K. Mass spectrometric analysis of peptides associated with the human class I MHC molecules HLA-A2.1 and HLA-B7 and identification of structural features that determine binding. Chem Immunol. 1993;57:39–62. [PubMed] [Google Scholar]
  14. Gaugler B., Van den Eynde B., van der Bruggen P., Romero P., Gaforio J. J., De Plaen E., Lethé B., Brasseur F., Boon T. Human gene MAGE-3 codes for an antigen recognized on a melanoma by autologous cytolytic T lymphocytes. J Exp Med. 1994 Mar 1;179(3):921–930. doi: 10.1084/jem.179.3.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Germain R. N., Margulies D. H. The biochemistry and cell biology of antigen processing and presentation. Annu Rev Immunol. 1993;11:403–450. doi: 10.1146/annurev.iy.11.040193.002155. [DOI] [PubMed] [Google Scholar]
  16. Hammond S. A., Bollinger R. C., Tobery T. W., Silliciano R. F. Transporter-independent processing of HIV-1 envelope protein for recognition by CD8+ T cells. Nature. 1993 Jul 8;364(6433):158–161. doi: 10.1038/364158a0. [DOI] [PubMed] [Google Scholar]
  17. Hearing V. J., Jiménez M. Analysis of mammalian pigmentation at the molecular level. Pigment Cell Res. 1989 Mar-Apr;2(2):75–85. doi: 10.1111/j.1600-0749.1989.tb00166.x. [DOI] [PubMed] [Google Scholar]
  18. Hearing V. J., Jiménez M. Mammalian tyrosinase--the critical regulatory control point in melanocyte pigmentation. Int J Biochem. 1987;19(12):1141–1147. doi: 10.1016/0020-711x(87)90095-4. [DOI] [PubMed] [Google Scholar]
  19. Henderson R. A., Cox A. L., Sakaguchi K., Appella E., Shabanowitz J., Hunt D. F., Engelhard V. H. Direct identification of an endogenous peptide recognized by multiple HLA-A2.1-specific cytotoxic T cells. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10275–10279. doi: 10.1073/pnas.90.21.10275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Henderson R. A., Michel H., Sakaguchi K., Shabanowitz J., Appella E., Hunt D. F., Engelhard V. H. HLA-A2.1-associated peptides from a mutant cell line: a second pathway of antigen presentation. Science. 1992 Mar 6;255(5049):1264–1266. doi: 10.1126/science.1546329. [DOI] [PubMed] [Google Scholar]
  21. Huczko E. L., Bodnar W. M., Benjamin D., Sakaguchi K., Zhu N. Z., Shabanowitz J., Henderson R. A., Appella E., Hunt D. F., Engelhard V. H. Characteristics of endogenous peptides eluted from the class I MHC molecule HLA-B7 determined by mass spectrometry and computer modeling. J Immunol. 1993 Sep 1;151(5):2572–2587. [PubMed] [Google Scholar]
  22. Hunt D. F., Henderson R. A., Shabanowitz J., Sakaguchi K., Michel H., Sevilir N., Cox A. L., Appella E., Engelhard V. H. Characterization of peptides bound to the class I MHC molecule HLA-A2.1 by mass spectrometry. Science. 1992 Mar 6;255(5049):1261–1263. doi: 10.1126/science.1546328. [DOI] [PubMed] [Google Scholar]
  23. Jerome K. R., Domenech N., Finn O. J. Tumor-specific cytotoxic T cell clones from patients with breast and pancreatic adenocarcinoma recognize EBV-immortalized B cells transfected with polymorphic epithelial mucin complementary DNA. J Immunol. 1993 Aug 1;151(3):1654–1662. [PubMed] [Google Scholar]
  24. Kaartinen V., Mononen T., Laatikainen R., Mononen I. Substrate specificity and reaction mechanism of human glycoasparaginase. The N-glycosidic linkage of various glycoasparagines is cleaved through a reaction mechanism similar to L-asparaginase. J Biol Chem. 1992 Apr 5;267(10):6855–6858. [PubMed] [Google Scholar]
  25. Kawakami Y., Eliyahu S., Delgado C. H., Robbins P. F., Rivoltini L., Topalian S. L., Miki T., Rosenberg S. A. Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3515–3519. doi: 10.1073/pnas.91.9.3515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kawakami Y., Eliyahu S., Delgado C. H., Robbins P. F., Sakaguchi K., Appella E., Yannelli J. R., Adema G. J., Miki T., Rosenberg S. A. Identification of a human melanoma antigen recognized by tumor-infiltrating lymphocytes associated with in vivo tumor rejection. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6458–6462. doi: 10.1073/pnas.91.14.6458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kawakami Y., Eliyahu S., Sakaguchi K., Robbins P. F., Rivoltini L., Yannelli J. R., Appella E., Rosenberg S. A. Identification of the immunodominant peptides of the MART-1 human melanoma antigen recognized by the majority of HLA-A2-restricted tumor infiltrating lymphocytes. J Exp Med. 1994 Jul 1;180(1):347–352. doi: 10.1084/jem.180.1.347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kwon B. S., Haq A. K., Pomerantz S. H., Halaban R. Isolation and sequence of a cDNA clone for human tyrosinase that maps at the mouse c-albino locus. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7473–7477. doi: 10.1073/pnas.84.21.7473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Michaëlsson E., Malmström V., Reis S., Engström A., Burkhardt H., Holmdahl R. T cell recognition of carbohydrates on type II collagen. J Exp Med. 1994 Aug 1;180(2):745–749. doi: 10.1084/jem.180.2.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mononen I., Fisher K. J., Kaartinen V., Aronson N. N., Jr Aspartylglycosaminuria: protein chemistry and molecular biology of the most common lysosomal storage disorder of glycoprotein degradation. FASEB J. 1993 Oct;7(13):1247–1256. doi: 10.1096/fasebj.7.13.8405810. [DOI] [PubMed] [Google Scholar]
  31. Moore M. W., Carbone F. R., Bevan M. J. Introduction of soluble protein into the class I pathway of antigen processing and presentation. Cell. 1988 Sep 9;54(6):777–785. doi: 10.1016/s0092-8674(88)91043-4. [DOI] [PubMed] [Google Scholar]
  32. Morrison L. A., Lukacher A. E., Braciale V. L., Fan D. P., Braciale T. J. Differences in antigen presentation to MHC class I-and class II-restricted influenza virus-specific cytolytic T lymphocyte clones. J Exp Med. 1986 Apr 1;163(4):903–921. doi: 10.1084/jem.163.4.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Oetting W. S., Handoko H. Y., Mentink M. M., Paller A. S., White J. G., King R. A. Molecular analysis of an extended family with type IA (tyrosinase-negative) oculocutaneous albinism. J Invest Dermatol. 1991 Jul;97(1):15–19. doi: 10.1111/1523-1747.ep12477808. [DOI] [PubMed] [Google Scholar]
  34. Ohkura T., Yamashita K., Mishima Y., Kobata A. Purification of hamster melanoma tyrosinases and structural studies of their asparagine-linked sugar chains. Arch Biochem Biophys. 1984 Nov 15;235(1):63–77. doi: 10.1016/0003-9861(84)90255-8. [DOI] [PubMed] [Google Scholar]
  35. Ruppert J., Sidney J., Celis E., Kubo R. T., Grey H. M., Sette A. Prominent role of secondary anchor residues in peptide binding to HLA-A2.1 molecules. Cell. 1993 Sep 10;74(5):929–937. doi: 10.1016/0092-8674(93)90472-3. [DOI] [PubMed] [Google Scholar]
  36. Salter R. D., Howell D. N., Cresswell P. Genes regulating HLA class I antigen expression in T-B lymphoblast hybrids. Immunogenetics. 1985;21(3):235–246. doi: 10.1007/BF00375376. [DOI] [PubMed] [Google Scholar]
  37. Slingluff C. L., Jr, Cox A. L., Henderson R. A., Hunt D. F., Engelhard V. H. Recognition of human melanoma cells by HLA-A2.1-restricted cytotoxic T lymphocytes is mediated by at least six shared peptide epitopes. J Immunol. 1993 Apr 1;150(7):2955–2963. [PubMed] [Google Scholar]
  38. Suzuki T., Seko A., Kitajima K., Inoue Y., Inoue S. Identification of peptide:N-glycanase activity in mammalian-derived cultured cells. Biochem Biophys Res Commun. 1993 Aug 16;194(3):1124–1130. doi: 10.1006/bbrc.1993.1938. [DOI] [PubMed] [Google Scholar]
  39. Suzuki T., Seko A., Kitajima K., Inoue Y., Inoue S. Purification and enzymatic properties of peptide:N-glycanase from C3H mouse-derived L-929 fibroblast cells. Possible widespread occurrence of post-translational remodification of proteins by N-deglycosylation. J Biol Chem. 1994 Jul 1;269(26):17611–17618. [PubMed] [Google Scholar]
  40. Townsend A., Bodmer H. Antigen recognition by class I-restricted T lymphocytes. Annu Rev Immunol. 1989;7:601–624. doi: 10.1146/annurev.iy.07.040189.003125. [DOI] [PubMed] [Google Scholar]
  41. Traversari C., van der Bruggen P., Luescher I. F., Lurquin C., Chomez P., Van Pel A., De Plaen E., Amar-Costesec A., Boon T. A nonapeptide encoded by human gene MAGE-1 is recognized on HLA-A1 by cytolytic T lymphocytes directed against tumor antigen MZ2-E. J Exp Med. 1992 Nov 1;176(5):1453–1457. doi: 10.1084/jem.176.5.1453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Utz U., Koenig S., Coligan J. E., Biddison W. E. Presentation of three different viral peptides, HTLV-1 Tax, HCMV gB, and influenza virus M1, is determined by common structural features of the HLA-A2.1 molecule. J Immunol. 1992 Jul 1;149(1):214–221. [PubMed] [Google Scholar]
  43. Wright H. T. Nonenzymatic deamidation of asparaginyl and glutaminyl residues in proteins. Crit Rev Biochem Mol Biol. 1991;26(1):1–52. doi: 10.3109/10409239109081719. [DOI] [PubMed] [Google Scholar]
  44. Wölfel T., Van Pel A., Brichard V., Schneider J., Seliger B., Meyer zum Büschenfelde K. H., Boon T. Two tyrosinase nonapeptides recognized on HLA-A2 melanomas by autologous cytolytic T lymphocytes. Eur J Immunol. 1994 Mar;24(3):759–764. doi: 10.1002/eji.1830240340. [DOI] [PubMed] [Google Scholar]
  45. Yewdell J. W., Bennink J. R. Cell biology of antigen processing and presentation to major histocompatibility complex class I molecule-restricted T lymphocytes. Adv Immunol. 1992;52:1–123. doi: 10.1016/s0065-2776(08)60875-5. [DOI] [PubMed] [Google Scholar]
  46. den Haan J. M., Sherman N. E., Blokland E., Huczko E., Koning F., Drijfhout J. W., Skipper J., Shabanowitz J., Hunt D. F., Engelhard V. H. Identification of a graft versus host disease-associated human minor histocompatibility antigen. Science. 1995 Jun 9;268(5216):1476–1480. doi: 10.1126/science.7539551. [DOI] [PubMed] [Google Scholar]
  47. van Binnendijk R. S., van Baalen C. A., Poelen M. C., de Vries P., Boes J., Cerundolo V., Osterhaus A. D., UytdeHaag F. G. Measles virus transmembrane fusion protein synthesized de novo or presented in immunostimulating complexes is endogenously processed for HLA class I- and class II-restricted cytotoxic T cell recognition. J Exp Med. 1992 Jul 1;176(1):119–128. doi: 10.1084/jem.176.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. van der Bruggen P., Traversari C., Chomez P., Lurquin C., De Plaen E., Van den Eynde B., Knuth A., Boon T. A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science. 1991 Dec 13;254(5038):1643–1647. doi: 10.1126/science.1840703. [DOI] [PubMed] [Google Scholar]

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