Interaction of HLA-E with peptides and the peptide transporter in vitro: implications for its function in antigen presentation.
The assembly of MHC Ia molecules in the endoplasmic reticulum requires the presence of peptide ligands and beta2m and is facilitated by chaperones in an ordered sequence of molecular interactions. A crucial step in this process is the interaction of the class I alpha-chain/beta2m dimer with TAP, which is believed to ensure effective peptide loading of the empty class I molecule. We have previously demonstrated impaired intracellular transport of the class Ib molecule HLA-E in mouse myeloma cells cotransfected with the genes for HLA-E and human beta2m, which is most likely attributable to inefficient intracellular peptide loading of the HLA-E molecule. We therefore analyzed the ability of HLA-E in the transfectant cell line to bind synthetic peptides by means of their ability to enhance cell surface expression of HLA-E. Peptide binding was confirmed by testing the effect on the thermostability of soluble empty HLA-E/human beta2m dimers. Two viral peptides binding to HLA-E were thus identified, for which the exact positioning of the N terminus appeared critical for binding, whereas the contribution of the length of the C terminus seemed to be minor, allowing peptides as short as seven amino acids and up to 16 amino acids to exhibit considerable binding activity. Furthermore, we demonstrate that HLA-E interacts with TAP and that this interaction can be prolonged by the proteasome inhibitor N-acetyl-L-leucyl-L-leucyl-L-norleucinal, which reduces the intracellular peptide pool. The presented data indicate that HLA-E is capable of presenting peptide ligands similar to the repertoire of HLA class Ia molecules.
Ulbrecht
M
M
Institut für Anthropologie und Humangenetik, Ludwig-Maximilians-Universität München, Munich, Germany.
Modrow
S
S
Srivastava
R
R
Peterson
P A
PA
Weiss
E H
EH
D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation; D017920:Antiporters; D002352:Carrier Proteins / Q000276:immunology*; D005434:Flow Cytometry; D006680:HLA Antigens / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006801:Humans; D007136:Immunoglobulins; D026901:Membrane Transport Proteins; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D011233:Precipitin Tests; D014407:Tumor Cells, Cultured; D000097625:HLA-E Antigens
D017920:Antiporters; D002352:Carrier Proteins; D006680:HLA Antigens; D015395:Histocompatibility Antigens Class I; D007136:Immunoglobulins; D026901:Membrane Transport Proteins; D010455:Peptides; D000097625:HLA-E Antigens; C107762:tapasin