title: Melanoma cells present high levels of HLA-A2-tyrosinase in association with instability and aberrant intracellular processing of tyrosinase.
abstract: Short-lived protein translation products are proposed to be a major source of substrates for major histocompatibility complex (MHC) class I antigen processing and presentation; however, a direct link between protein stability and the presentation level of MHC class I-peptide complexes has not been made. We have recently discovered that the peptide Tyr((369-377)) , derived from the tyrosinase protein is highly presented by HLA-A2 on the surface of melanoma cells. To examine the molecular mechanisms responsible for this presentation, we compared characteristics of tyrosinase in melanoma cells lines that present high or low levels of HLA-A2-Tyr((369-377))  complexes. We found no correlation between mRNA levels and the levels of HLA-A2-Tyr((369-377))  presentation. Co-localization experiments revealed that, in cell lines presenting low levels of HLA-A2-Tyr((369-377))  complexes, tyrosinase co-localizes with LAMP-1, a melanosome marker, whereas in cell lines presenting high HLA-A2-Tyr((369-377))  levels, tyrosinase localizes to the endoplasmic reticulum. We also observed differences in tyrosinase molecular weight and glycosylation composition as well as major differences in protein stability (t(1/2) ). By stabilizing the tyrosinase protein, we observed a dramatic decrease in HLA-A2-tyrosinase presentation. Our findings suggest that aberrant processing and instability of tyrosinase are responsible for the high presentation of HLA-A2-Tyr((369-377))  complexes and thus shed new light on the relationship between intracellular processing, stability of proteins, and MHC-restricted peptide presentation.
authors.affiliation: Faculty of Biology, Technion-Israel Institute of Technology, Haifa, Israel.
authors.forename: Yael
authors.initials: Y
authors.lastname: Michaeli
authors.forename: Keren
authors.initials: K
authors.lastname: Sinik
authors.forename: Maya
authors.initials: M
authors.lastname: Haus-Cohen
authors.forename: Yoram
authors.initials: Y
authors.lastname: Reiter
mesh_terms: D017951:Antigen Presentation; D045744:Cell Line, Tumor; D006031:Glycosylation; D015789:HLA-A2 Antigen / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D051907:Lysosomal Membrane Proteins / Q000276:immunology / Q000378:metabolism; D008545:Melanoma / Q000276:immunology* / Q000378:metabolism; D020460:Melanosomes / Q000276:immunology / Q000378:metabolism; D014442:Monophenol Monooxygenase / Q000276:immunology* / Q000378:metabolism; D009363:Neoplasm Proteins / Q000276:immunology* / Q000378:metabolism; D010455:Peptides / Q000276:immunology / Q000378:metabolism; D055550:Protein Stability; D012333:RNA, Messenger / Q000276:immunology / Q000378:metabolism; D052118:Lysosomal-Associated Membrane Protein 1
chemical_list: D015789:HLA-A2 Antigen; D051907:Lysosomal Membrane Proteins; D014442:Monophenol Monooxygenase; D009363:Neoplasm Proteins; D010455:Peptides; D012333:RNA, Messenger; C489116:LAMP1 protein, human; D052118:Lysosomal-Associated Membrane Protein 1