Identification and characterization of three distinct families of glycoprotein complexes in the envelopes of human cytomegalovirus.
Several disulfide-linked glycoprotein complexes were identified in the envelope of human cytomegalovirus (HCMV). These glycoprotein complexes were fractionated by rate-zonal centrifugation in sucrose density gradients in the presence of detergents. Fractionated glycoproteins and complexes were immunoprecipitated with three different monoclonal antibodies specific for HCMV glycoproteins and a rabbit polyclonal antiserum prepared against detergent-extracted virion and dense-body envelope glycoproteins. Three distinct families of disulfide-linked glycoprotein complexes were observed and designated glycoprotein complex gcI, gcII, and gcIII. The gcI family, recognized by monoclonal antibody 41C2 under nonreducing conditions, consisted of three complexes with approximate molecular masses of 250 to 300, 190, and 160 kilodaltons (kDa). These complexes consistently sediment more rapidly than other HCMV glycoproteins or complexes in sucrose density gradients. Upon reduction of the gcI family, two size classes of glycoproteins with average molecular masses of 93 to 130 and 55 kDa were observed. The gcII family was recognized by monoclonal antibody 9E10. Under nonreducing conditions, as many as six electrophoretic forms were observed for gcII. When reduced, the major component of the gcII family was a heterogeneous glycoprotein designated gp47-52. The gcIII family was recognized by monoclonal antibody 1G6. It consisted of a complex of approximately 240 kDa without reduction of disulfide bonds. When reduced, two glycoprotein size classes with average molecular masses of 145 and 86 kDa were observed. Polyclonal antiserum R-7 reacted strongly with the gcI and gcIII families, but weakly with the gcII family.
Gretch
D R
DR
Department of Microbiology, University of Iowa, Iowa City 52242.
Kari
B
B
Rasmussen
L
L
Gehrz
R C
RC
Stinski
M F
MF
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002499:Centrifugation, Density Gradient; D003587:Cytomegalovirus / Q000032:analysis* / Q000276:immunology; D006023:Glycoproteins / Q000145:classification* / Q000276:immunology / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000145:classification* / Q000276:immunology / Q000302:isolation & purification; D014771:Virion / Q000032:analysis
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006023:Glycoproteins; D014759:Viral Envelope Proteins