Expression, Purification, and Crystallization of HSV-1 Glycoproteins for Structure Determination

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2060; p. 377
• Main Authors: White, Ellen M, Stampfer, Samuel D, Heldwein, Ekaterina E
• Format: Journal Article
• Language: English
• Published: United States 01.01.2020
• Subjects: Animals; Crystallography, X-Ray; Gene Expression; Glycoproteins - biosynthesis; Glycoproteins - chemistry; Glycoproteins - genetics; Glycoproteins - isolation & purification; Herpesvirus 1, Human - chemistry; Herpesvirus 1, Human - genetics; Herpesvirus 1, Human - metabolism; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Sf9 Cells; Spodoptera; Viral Envelope Proteins - biosynthesis; Viral Envelope Proteins - chemistry; Viral Envelope Proteins - genetics; Viral Envelope Proteins - isolation & purification; Viral entry; Glycoproteins; Protein purification; Crystallography; Herpes simplex viruses; Ectodomain
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-9814-2_23

Herpes simplex viruses utilize glycoproteins displayed on the viral envelope to perform a variety of functions in the viral infectious cycle. Structural and functional studies of these viral glycoproteins can benefit from biochemical, biophysical, and structural analysis of purified proteins. Here, we describe a general protocol for expression and purification of viral glycoproteins from insect cells based on those developed for the HSV-1 gB and HSV-2 gH/gL ectodomains as well as the protocol for crystallization of these glycoproteins. This protocol can be used for generating milligram amounts of wild-type (WT) or mutant gB and gH/gL ectodomains or can be adapted to produce purified ectodomains of glycoproteins from HSV or other herpesviruses for biochemical and structural studies.