J. Virol., 11 1996, 7379-7387, Vol 70, No. 11
Copyright © 1996, American Society for Microbiology

Disulfide bonds of herpes simplex virus type 2 glycoprotein gB

N Norais, D Tang, S Kaur, SH Chamberlain, FR Masiarz, RL Burke and F Marcus
Chiron Corporation, Emeryville, California 94608, USA.

Glycoprotein B (gB) is the most highly conserved envelope glycoprotein of herpesviruses. The gB protein is required for virus infectivity and cell penetration. Recombinant forms of gB being used for the development of subunit vaccines are able to induce virus-neutralizing antibodies and protective efficacy in animal models. To gain structural information about the protein, we have determined the location of the disulfide bonds of a 696-amino-acid residue truncated, recombinant form of herpes simplex virus type 2 glycoprotein gB (HSV gB2t) produced by expression in Chinese hamster ovary cells. The purified protein, which contains virtually the entire extracellular domain of herpes simplex virus type 2 gB, was digested with trypsin under nonreducing conditions, and peptides were isolated by reversed-phase high- performance liquid chromatography (HPLC). The peptides were characterized by using mass spectrometry and amino acid sequence analysis. The conditions of cleavage (4 M urea, pH 7) induced partial carbamylation of the N termini of the peptides, and each disulfide peptide was found with two or three different HPLC retention times (peptides with and without carbamylation of either one or both N termini). The 10 cysteines of the molecule were found to be involved in disulfide bridges. These bonds were located between Cys-89 (C1) and Cys- 548 (C8), Cys-106 (C2) and Cys-504 (C7), Cys-180 (C3) and Cys-244 (C4), Cys-337 (C5) and Cys-385 (C6), and Cys-571 (C9) and Cys-608 (C10). These disulfide bonds are anticipated to be similar in the corresponding gBs from other herpesviruses because the 10 cysteines listed above are always conserved in the corresponding protein sequences.

This article has been cited by other articles:

• Heldwein, E. E., Lou, H., Bender, F. C., Cohen, G. H., Eisenberg, R. J., Harrison, S. C. (2006). Crystal structure of glycoprotein B from herpes simplex virus 1.. Science 313: 217-220 [Abstract] [Full Text]  
• Britt, W. J., Jarvis, M. A., Drummond, D. D., Mach, M. (2005). Antigenic Domain 1 Is Required for Oligomerization of Human Cytomegalovirus Glycoprotein B. J. Virol. 79: 4066-4079 [Abstract] [Full Text]  
• Banks, M., King, D. P., Daniells, C., Stagg, D. A., Gavier-Widen, D. (2002). Partial characterization of a novel gammaherpesvirus isolated from a European badger (Meles meles). J. Gen. Virol. 83: 1325-1330 [Abstract] [Full Text]  
• Lopper, M., Compton, T. (2002). Disulfide Bond Configuration of Human Cytomegalovirus Glycoprotein B. J. Virol. 76: 6073-6082 [Abstract] [Full Text]  
• Speckner, A., Kropff, B., Knör, S., Mach, M. (2000). The antigenic domain 1 of human cytomegalovirus glycoprotein B contains an intramolecular disulphide bond. J. Gen. Virol. 81: 2659-2663 [Abstract] [Full Text]  
• Trybala, E., Liljeqvist, J.-A., Svennerholm, B., Bergström, T. (2000). Herpes Simplex Virus Types 1 and 2 Differ in Their Interaction with Heparan Sulfate. J. Virol. 74: 9106-9114 [Abstract] [Full Text]  
• Peng, T., Ponce de Leon, M., Novotny, M. J., Jiang, H., Lambris, J. D., Dubin, G., Spear, P. G., Cohen, G. H., Eisenberg, R. J. (1998). Structural and Antigenic Analysis of a Truncated Form of the Herpes Simplex Virus Glycoprotein gH-gL Complex. J. Virol. 72: 6092-6103 [Abstract] [Full Text]  
• Laquerre, S., Anderson, D. B., Argnani, R., Glorioso, J. C. (1998). Herpes Simplex Virus Type 1 Glycoprotein B Requires a Cysteine Residue at Position 633 for Folding, Processing, and Incorporation into Mature Infectious Virus Particles. J. Virol. 72: 4940-4949 [Abstract] [Full Text]