Dissecting the structural determinants of the interaction between the human cytomegalovirus UL18 protein and the CD85j immune receptor.
UL18 is a glycoprotein encoded by the human cytomegalovirus genome and is thought to play a pivotal role during human cytomegalovirus infection, although its exact function is still a matter of debate. UL18 shares structural similarity with MHC class I and binds the receptor CD85j on immune cells. Besides UL18, CD85j binds MHC class I molecules. The binding properties of CD85j to MHC class I molecules have been thoroughly studied. Conversely, very little information is available on the CD85j/UL18 complex, namely that UL18 binds CD85j through its alpha3 domain with an affinity that is approximately 1000-fold higher than the MHC class I affinity for CD85j. Deeper knowledge of features of the UL18/CD85j complex would help to disclose the function of UL18 when it binds to CD85j. In this study we first demonstrated that the UL18alpha3 domain is not sufficient per se for binding and that beta2-microglobulin is necessary for UL18-CD85j interaction. We then dissected structural determinants of binding UL18 to CD85j. To this end, we constructed a three-dimensional model of the complex. The model was used to design mutants in selected regions of the putative interaction interface, the effects of which were measured on binding. Six regions in both the alpha2 and alpha3 domains and specific amino acids within them were identified that are potentially involved in the UL18-CD85j interaction. The higher affinity of UL18 to CD85j, compared with MHC class I, seems to be due not to additional interaction regions but to an overall better fit of the two molecules.
Occhino
Marzia
M
Department of Experimental Medicine, University of Genoa, Genoa, Italy.
Ghiotto
Fabio
F
Soro
Simonetta
S
Mortarino
Mimosa
M
Bosi
Stefania
S
Maffei
Massimo
M
Bruno
Silvia
S
Nardini
Marco
M
Figini
Mariangela
M
Tramontano
Anna
A
Ciccone
Ermanno
E
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000276:immunology*; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D017124:Conserved Sequence; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins; D006801:Humans; D000075362:Leukocyte Immunoglobulin-like Receptor B1; D008969:Molecular Sequence Data; D010940:Plant Proteins; D011971:Receptors, Immunologic / Q000276:immunology*
D015703:Antigens, CD; D036022:Capsid Proteins; D004268:DNA-Binding Proteins; C105299:LILRB1 protein, human; D000075362:Leukocyte Immunoglobulin-like Receptor B1; D010940:Plant Proteins; D011971:Receptors, Immunologic; C079750:VP23 protein, Human herpesvirus 1; C068831:drp90 protein, Glycine max