A cell free protein fragment complementation assay for monitoring the core interaction of the human cytomegalovirus nuclear egress complex

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• 作者：Margit Schnee¹ ; ² ; Felicia M. Wagner² ; Ulrich H. Koszinowski ; Zsolt Ruzsics ; ^{ruzsics@lmb.uni-muenchen.de}
• 关键词：HCMV ; human cytomegalovirus ; NEC ; nuclear egress complex ; PPI ; Protein&ndash ; protein interactions ; CR1-4 ; conserved region one to four ; Bla ; 尾-lactamase ; PCA ; protein fragment complementation assay
• 刊名：Antiviral Research
• 出版年：2012
• 期刊代码：7_01663542
• 类别：imm
• 出版时间：July, 2012
• 卷：95
• 期：1
• 页码：12-18
• 文件大小：416 K

摘要

Certain viral protein-protein interactions provide attractive targets for antiviral drug development. Recently, we described a 尾-lactamase based protein fragment complementation assay (PCA) to study the core interaction of the nuclear egress complex (NEC) of different herpesviruses in cells. Now, to have a cell free assay for inhibitor screens, we expressed split 尾-lactamase tagged interaction domains of the viral pUL50 and pUL53 proteins representing the NEC of human cytomegalovirus (HCMV) in bacteria. After validation and basic characterization of this NEC-PCA, we tested peptide inhibitors of the pUL50-pUL53 complex. We show that peptides resembling sequences of the first conserved region of pUL53 can inhibit the NEC-PCA. This, on one hand, indicated that the core interaction in the HCMV NEC is mediated by a linear motif. On the other hand it proved that this new pUL50-pUL53 interaction assay allows a simple cell free test for small molecular inhibitors.