Related Experiment Video
Updated: Jan 5, 2026
08:58
Characterization of Glycoproteins with the Immunoglobulin Fold by X-Ray Crystallography and Biophysical Techniques
Published on: July 5, 2018
13.1K
Expression, Purification, and Crystallization of HSV-1 Glycoproteins for Structure Determination
Ellen M White1, Samuel D Stampfer1, Ekaterina E Heldwein2
1Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|October 17, 2019
Summary
This study presents a general protocol for expressing and purifying herpes simplex virus (HSV) glycoproteins from insect cells. This method facilitates biochemical and structural studies of viral glycoproteins essential for understanding HSV infection.
Area of Science:
- Virology
- Structural Biology
- Protein Biochemistry
Background:
- Herpes simplex viruses (HSV) use envelope glycoproteins for critical functions during infection.
- Understanding these glycoproteins requires purified proteins for biochemical, biophysical, and structural analysis.
Purpose of the Study:
- To describe a generalizable protocol for the expression and purification of viral glycoproteins from insect cells.
- To enable structural and functional studies of HSV glycoproteins.
Main Methods:
- Expression of wild-type or mutant glycoprotein ectodomains (e.g., HSV-1 gB, HSV-2 gH/gL) in insect cells.
- Purification of milligram quantities of these glycoproteins.
- Adaptation of protocols for crystallization.
Main Results:
- A versatile protocol for producing purified viral glycoprotein ectodomains is established.
- The protocol is based on successful methods for HSV-1 gB and HSV-2 gH/gL.
- The method yields sufficient protein for detailed biochemical and structural investigations.
Conclusions:
- The described protocol provides a reliable method for obtaining purified herpesvirus glycoproteins.
- This facilitates further research into the structure and function of these essential viral components.
- The protocol can be adapted for glycoproteins from HSV and other herpesviruses.
