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

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Herpes Simplex Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2060))

Abstract

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.

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Acknowledgments

We acknowledge the contributions of the laboratory of Roselyn Eisenberg and Gary Cohen toward the development of the initial purification protocols of HSV-1 gB730 and HSV-2 gH803/gL produced using recombinant baculoviruses. We also thank Tirumala K. Chowdary and Sapna Sharma for their work in establishing and optimizing these protocols in our laboratory. Finally, we thank past and present members of the Heldwein lab for helpful advice and discussions.

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Authors and Affiliations

  1. Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA, USA

    Ellen M. White, Samuel D. Stampfer & Ekaterina E. Heldwein

Authors
  1. Ellen M. White
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  2. Samuel D. Stampfer
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  3. Ekaterina E. Heldwein
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Corresponding author

Correspondence to Ekaterina E. Heldwein .

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Editors and Affiliations

  1. Department of Biomedical Sciences, Macquarie University, Sydney, NSW, Australia

    Russell J. Diefenbach

  2. Institute of Virology, University of Zurich, Zürich, Switzerland

    Cornel Fraefel

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White, E.M., Stampfer, S.D., Heldwein, E.E. (2020). Expression, Purification, and Crystallization of HSV-1 Glycoproteins for Structure Determination. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus . Methods in Molecular Biology, vol 2060. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9814-2_23

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  • DOI: https://doi.org/10.1007/978-1-4939-9814-2_23

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9813-5

  • Online ISBN: 978-1-4939-9814-2

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