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

  • Tri Komala Sari
  • Katrina A. Gianopulos
  • Anthony V. NicolaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2060)


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.

Key words

Immunoassay Dot blot Antibodies Nitrocellulose membrane Virus entry Glycoproteins Herpesvirus Herpes simplex virus Conformational change Membrane fusion Low pH 



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


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Tri Komala Sari
    • 1
  • Katrina A. Gianopulos
    • 1
    • 2
  • Anthony V. Nicola
    • 1
    • 2
    Email author
  1. 1.Department of Veterinary Microbiology and Pathology, College of Veterinary MedicineWashington State UniversityPullmanUSA
  2. 2.School of Molecular Biosciences, College of Veterinary MedicineWashington State UniversityPullmanUSA

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