Orientation of the cleavage site of the herpes simplex virus glycoprotein G-2.
During the synthesis of glycoprotein G-2 (gG-2) of herpes simplex virus type 2, the 104,000-Da gG-2 precursor (104K precursor) is cleaved to generate the 72K and the 31K intermediates. The 72K product is processed to generate the mature gG-2 (molecular mass, 108,000 Da), while the 31K product is additionally processed and secreted into the extracellular medium as the 34K component (H. K. Su, R. Eberle, and R. J. Courtney, J. Virol. 61:1735-1737, 1987). In this study, the orientations of the 31K and 72K products on the 104K precursor were determined by using two antipeptide sera produced in rabbits and a monoclonal antibody, 13 alpha C6, directed against gG-2. The sera prepared against synthetic peptides corresponding to the terminal amino acid residues 67 to 78 and an internal peptide at amino acids 247 to 260 of gG-2 recognized the 104K precursor and the 31K cleavage product but not the 72K intermediate. In contrast, 13 alpha C6 detected the 72K cleavage product and the uncleaved precursor but not the 31K cleavage component. The epitope recognized by 13 alpha C6 was mapped within amino acids 486 to 566. These results suggest that the 31K cleavage product is derived from the amino-terminal portion of the 104K precursor molecule and that the 72K intermediate is derived from the carboxyl terminus. In support of our model described above for the synthesis of gG-2, antibodies recognizing either of the cleavage products reacted with the uncleaved precursor but not with the other cleavage product. By using partial endo-beta-N-acetylglucosaminidase H analysis, two N-linked glycosylation sites were found on each of the cleavage products. The distribution of the N-linked glycosylation sites and the reactivities of the antipeptide sera allowed the cleavage region on the precursor to be mapped to within amino acids 260 to 437.
Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, Hershey 17033.
H K
HK
Su
J D
JD
Fetherston
M E
ME
Smith
R J
RJ
Courtney
D000911:Antibodies, Monoclonal; D004252:DNA Mutational Analysis; D006031:Glycosylation; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology; D011499:Protein Processing, Post-Translational; D018139:Simplexvirus / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D014759:Viral Envelope Proteins; C045464:glycoprotein G, herpes simplex virus type 2; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase