Avitabile Elisa, Forghieri Cristina, Campadelli-Fiume Gabriella
Department of Experimental Pathology, Section on Microbiology and Virology, University of Bologna, Via San Giacomo 12, 40126 Bologna, Italy.
J Virol. 2007 Oct;81(20):11532-7. doi: 10.1128/JVI.01343-07. Epub 2007 Aug 1.
The interactions between herpes simplex virus gD and its nectin1 receptor or between gD, gB, and gH were analyzed by complementation of the N and C portions of split enhanced green fluorescent protein (EGFP) fused to the glycoproteins. The gD(N)-Nect(C) complex was readily detected; the gD(N)-gC(C) complex was undetectable, highlighting the specificity of the assay. Split EGFP complementation was detected between proteins designated gD(N)+gH(C), gD(N)+gB(C), and gH(N)+gB(C)+wtgD (gB was deleted of endocytosis motifs), both in cells transfected with two-tree glycoproteins and in syncytia. The in situ assay provides evidence that gD interacts with gH and gB independently of each other and supports a model whereby gH and gB in complex exert their activities to gD.
通过将与糖蛋白融合的分裂增强型绿色荧光蛋白（EGFP）的N端和C端进行互补，分析了单纯疱疹病毒gD与其nectin1受体之间或gD、gB和gH之间的相互作用。很容易检测到gD(N)-Nect(C)复合物；而gD(N)-gC(C)复合物则检测不到，这突出了该检测方法的特异性。在转染了两种糖蛋白的细胞以及多核细胞中，在指定的gD(N)+gH(C)、gD(N)+gB(C)和gH(N)+gB(C)+wtgD（gB缺失内吞基序）的蛋白质之间均检测到了分裂EGFP互补。原位检测提供了证据，表明gD与gH和gB相互独立地相互作用，并支持一种模型，即gH和gB形成的复合物对gD发挥其活性作用。
