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Conformational Change in Herpes Simplex Virus Entry Glycoproteins Detected by Dot Blot

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Herpes Simplex Virus

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

Abstract

Conformational changes in viral membrane proteins drive membrane fusion, a critical step in virus entry and infection. Here we describe a simple and rapid virus blotting immunoassay to define conformational changes with a panel of monoclonal antibodies to distinct sites across a viral glycoprotein. This dot blot technique has been utilized to define low pH-triggered changes in the prefusion form of the herpesviral fusogen gB. At pH of <6.2 there are specific changes in herpes simplex virus 1 gB domains I and V. This corresponds broadly to host cell endosomal pH. Many of the identified changes are at least partially reversible. This method can be adapted to document changes in viral proteins that are not fusion proteins, including those induced by alternate triggers such as receptor-binding or protease cleavage.

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Acknowledgments

This work was supported by Public Health Service grants AI119159 and GM008336 from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA, USA

    Tri Komala Sari, Katrina A. Gianopulos & Anthony V. Nicola

  2. School of Molecular Biosciences, College of Veterinary Medicine, Washington State University, Pullman, WA, USA

    Katrina A. Gianopulos & Anthony V. Nicola

Authors
  1. Tri Komala Sari
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  2. Katrina A. Gianopulos
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  3. Anthony V. Nicola
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Corresponding author

Correspondence to Anthony V. Nicola .

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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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Sari, T.K., Gianopulos, K.A., Nicola, A.V. (2020). Conformational Change in Herpes Simplex Virus Entry Glycoproteins Detected by Dot Blot. 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_18

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  • DOI: https://doi.org/10.1007/978-1-4939-9814-2_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9813-5

  • Online ISBN: 978-1-4939-9814-2

  • eBook Packages: Springer Protocols

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