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The herpesviral ICP47 inhibits TAP function, thereby suppressing an adaptive immune response. Here, we report on a thermostable ICP47-TAP complex, generated by fusion of different ICP47 fragments. These fusion complexes allowed us to determine the direction and positioning in the central cavity of TAP. ICP47-TAP fusion complexes are arrested in a stable conformation, as demonstrated by MHC I surface expression, melting temperature, and the mutual exclusion of herpesviral TAP inhibitors. We unveiled a conserved region next to the active domain of ICP47 as essential for the complete stabilization of the TAP complex. Binding of the active domain of ICP47 arrests TAP in an open inward facing conformation rendering the complex inaccessible for other viral factors. Based on our findings, we propose a dual interaction mechanism for ICP47. A <jats:italic>per se<\/jats:italic> destabilizing active domain inhibits the function of TAP, whereas a conserved C-terminal region additionally stabilizes the transporter. These new insights into the ICP47 inhibition mechanism can be applied for future structural analyses of the TAP complex.<\/jats:p>","DOI":"10.1038\/srep36907","type":"journal-article","created":{"date-parts":[[2016,11,15]],"date-time":"2016-11-15T10:51:42Z","timestamp":1479207102000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["A dual inhibition mechanism of herpesviral ICP47 arresting a conformationally thermostable TAP complex"],"prefix":"10.1038","volume":"6","author":[{"given":"Valentina","family":"Herbring","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anja","family":"B\u00e4ucker","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simon","family":"Trowitzsch","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert","family":"Tamp\u00e9","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2016,11,15]]},"reference":[{"key":"BFsrep36907_CR1","doi-asserted-by":"publisher","first-page":"741","DOI":"10.1038\/348741a0","volume":"348","author":"J Trowsdale","year":"1990","unstructured":"Trowsdale, J. et al. Sequences encoded in the class II region of the MHC related to the \u2018ABC\u2019 superfamily of transporters. Nature 348, 741\u2013744 (1990).","journal-title":"Nature"},{"key":"BFsrep36907_CR2","doi-asserted-by":"publisher","first-page":"738","DOI":"10.1038\/348738a0","volume":"348","author":"EV Deverson","year":"1990","unstructured":"Deverson, E. V. et al. MHC class II region encoding proteins related to the multidrug resistance family of transmembrane transporters. Nature 348, 738\u2013741 (1990).","journal-title":"Nature"},{"key":"BFsrep36907_CR3","doi-asserted-by":"publisher","first-page":"10142","DOI":"10.1074\/jbc.M312816200","volume":"279","author":"J Koch","year":"2004","unstructured":"Koch, J., Guntrum, R., Heintke, S., Kyritsis, C. & Tamp\u00e9, R. Functional dissection of the transmembrane domains of the transporter associated with antigen processing (TAP). 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