Determination of the transmembrane topology of herpes simplex virus type 1 glycoprotein K.
Herpes simplex virus type 1 glycoprotein K (gK) plays an essential role in viral replication and cell fusion. gK is a very hydrophobic membrane protein that contains a signal sequence and several hydrophobic regions. It has been shown that mutations inducing cell fusion map to two distinct domains of gK, suggesting that these domains are functionally important. To understand the transmembrane topology of gK and the localization of these functional domains, we constructed a set of gK deletion, insertion, and truncation mutants and expressed these by in vitro translation in the presence of microsomal membranes. The transmembrane topology of gK was determined by examination of the post-translational processing and protease sensitivity of the mutant proteins. Our data demonstrate that gK contains three transmembrane domains (amino acids 125-139, 226-239, and 311-325). Another hydrophobic domain (amino acids 241-265), which is relatively less hydrophobic and much longer compared with the transmembrane sequences, is located in the extracellular loop. The analysis showed that the domains containing syncytial mutations are both ectodomains. They may interact with each other to form a complex tertiary structure that is critical for the biological function of gK.
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Department of Immunology and Microbiology, Wayne State University Medical School, Detroit, Michigan 48201, USA.
Holland
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004285:Dogs; D019286:Endopeptidase K / Q000378:metabolism; D006596:Hexosaminidases / Q000378:metabolism; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000737:chemistry; D008861:Microsomes / Q000737:chemistry; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics
D011994:Recombinant Proteins; C072448:UL53 protein, Human herpesvirus 1; D014764:Viral Proteins; D006596:Hexosaminidases; D019286:Endopeptidase K