The @entity4538 glycoprotein B gene (gB; gpUL55) was truncated at @entity535 692 and recombined into @entity20422 (baculovirus). Infection of insect cells with the recombinant baculovirus resulted in high-level expression and secretion of the truncated gB protein (gB-S) into the culture medium. Purification of gB-S by monoclonal antibody affinity chromatography yielded a protein of ca. 200 kDa. Characterization of the 200-kDa purification product indicated that the recombinant gB protein retained many structural and functional features of the viral gB. Comparison of electrophoretic migration patterns in reduced versus nonreduced protein samples and immune blotting analysis with antibodies specific for the @entity1472 or @entity1473 -terminus of gB demonstrated that the recombinant protein was composed of @entity1520 linked 69 kDa @entity1472 terminal and 35-kDa @entity1473 -terminal fragments. In addition, recognition of the 200-kDa gB-S by a conformational-dependent, oligomer-specific monoclonal antibody suggested that gB-S was properly folded and dimeric. Like the viral gB, gB-S had heparin binding ability. One heparin binding site was found to reside within the 35-kDa @entity1473 -terminal fragment (aa 492-692). Heparin binding was abolished when gB-S was denatured. These data suggest that gB contains a novel heparin binding motif that is at least partially conformational dependent.

Expression, purification, and characterization of a soluble form of XXXX glycoprotein B.

@entity1472 :: ('CHEBI:46882', 'Chemical') :: ['amino'] @entity20422 :: ('46015', 'Species') :: ['Autographa californica nuclear polyhedrosis virus'] @entity535 :: ('CHEBI:33704', 'Chemical') :: ['amino acid'] @entity4538 :: ('10359', 'Species') :: ['human cytomegalovirus'] @entity1520 :: ('CHEBI:48343', 'Chemical') :: ['disulfide'] @entity1473 :: ('CHEBI:46883', 'Chemical') :: ['carboxy']
@entity4538 :: (10359,Species) :: ['human cytomegalovirus']
