title: Identification of amino acids within the MHC molecule important for the interaction with the adenovirus protein E3/19K.
abstract: The E3/19K protein of human adenovirus type 2 binds to class I MHC Ags thereby interfering with their cell surface expression and Ag presentation function. Currently, it is unclear exactly which structure of MHC molecules is recognized by the E3/19K protein. We have previously demonstrated that the murine H-2Kd Ag is able to associate with E3/19K, whereas the allelic H-2Kk molecule is not. By using exon shuffling between Kd and Kk molecules, the alpha 1 and alpha 2 domains of MHC class I molecules were identified as essential structures for binding the viral protein. In this report, we have examined the contribution of individual amino acids within the alpha 2 domain of MHC for binding E3/19K. First, we show that within this domain the alpha-helical part is most important for the interaction with E3/19K. By using site-directed mutagenesis, Kd-specific amino acids were introduced into the alpha-helix of the alpha 2 domain of Kk. By using the expression of mutagenized proteins in E3/19K+ cells, we have identified Tyr 156 and Leu 180 as being essential for the association with the E3/19K protein. In addition, Kd residue Glu 163 seems to contribute to the complex formation. Furthermore, analysis of a panel of Kd/Dd recombinants indicates that a similar region in the Dd molecule, namely, the C-terminal half of the alpha 2 domain, affects binding to E3/19K. Combining these results with Ab binding data, we present two alternative models of how the adenovirus protein may bind to the alpha 1 and alpha 2 domains.
authors.affiliation: Hans Spemann Laboratory, Max Planck Institute for Immunobiology, Freiburg, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Feuerbach
authors.forename: S
authors.initials: S
authors.lastname: Etteldorf
authors.forename: C
authors.initials: C
authors.lastname: Ebenau-Jehle
authors.forename: J P
authors.initials: JP
authors.lastname: Abastado
authors.forename: D
authors.initials: D
authors.lastname: Madden
authors.forename: H G
authors.initials: HG
authors.lastname: Burgert
mesh_terms: D017503:Adenovirus E3 Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D000260:Adenoviruses, Human / Q000276:immunology*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D017931:DNA Primers / Q000737:chemistry; D006183:H-2 Antigens / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship
chemical_list: D017503:Adenovirus E3 Proteins; D017931:DNA Primers; D006183:H-2 Antigens; D011993:Recombinant Fusion Proteins