Characterization by capillary electrophoresis of the surface glycoproteins of ovine lentiviruses before and after treatment with glycosidic enzymes.
Ovine lentiviruses are a group of viruses that infect sheep and goats. These viruses contain a surface glycoprotein (SU) that is very similar among the viral strains. Sera from infected animals react equally well with SU from each strain. Monoclonal antibodies produced to SU can distinguish among some of the viral strains. In order to delineate these differences we treated SU from several viral strains with the glycosidic enzymes. These enzymes included a mixture of exoglycosidases, beta-N-acetyl glucosaminidase, neuraminidase and endoglycosidases D, F and H. After these treatments we observed changes in the reactivities of the monoclonal antibodies that were directed to SU. In order to characterize these changes on the surface epitopes, SU from the different viral strains were subjected to free zone capillary electrophoresis (CZE) using an 0.02 M phosphate buffer at pH 9.0 at a running voltage of 5 kV. Differences were readily seen between SU that had not been treated and SU that had been treated with the glycosidic enzymes. Each viral strain had a characteristic electropherogram. The electropherograms indicated that the heterogeneity of the charge on SU was increased after the enzyme treatments. From these results we have concluded that the carbohydrate moieties play an important role in contributing to the surface charge of SU. This charge affects the nature of its surface epitopes and has an impact on its biological function.
Schmerr
M J
MJ
National Animal Disease Center, USDA, Ames, IA 50010.
Goodwin
K R
KR
D000818:Animals; D004586:Electrophoresis / Q000379:methods*; D006026:Glycoside Hydrolases / Q000378:metabolism*; D016086:Lentivirus / Q000737:chemistry*; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011863:Radioimmunoassay; D012756:Sheep; D014764:Viral Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D008562:Membrane Glycoproteins; D014764:Viral Proteins; D006026:Glycoside Hydrolases