title: Disulfide bonds between two envelope proteins of lactate dehydrogenase-elevating virus are essential for viral infectivity.
abstract: Disulfide bonds were found to link the nonglycosylated envelope protein VP-2/M (19 kDa), encoded by open reading frame 6, and the major envelope glycoprotein VP-3 (25 to 42 kDa), encoded by open reading frame 5, of lactate dehydrogenase-elevating virus (LDV). The two proteins comigrated in a complex of 45 to 55 kDa when the virion proteins were electrophoresed under nonreducing conditions but dissociated under reducing conditions. Furthermore, VP-2/M was quantitatively precipitated along with VP-3 in this complex by three neutralizing monoclonal antibodies to VP-3. The infectivity of LDV was rapidly and irreversibly lost during incubation with 5 to 10 mM dithiothreitol (> 99% in 6 h at room temperature), which is known to reduce disulfide bonds. LDV inactivation correlated with dissociation of VP-2/M and VP-3. The results suggest that disulfide bonds between VP-2/M and VP-3 are important for LDV infectivity. Hydrophobic moment analyses of the predicted proteins suggest that VP-2/M and VP-3 both possess three adjacent transmembrane segments and only very short ectodomains (10 and 32 amino acids, respectively) with one and two cysteines, respectively. Inactivation of LDV by dithiothreitol and dissociation of the two envelope proteins were not associated with alterations in LDV's density or sedimentation coefficient.
authors.affiliation: Department of Microbiology, Medical School, University of Minnesota, Minneapolis 55455.
authors.forename: K S
authors.initials: KS
authors.lastname: Faaberg
authors.forename: C
authors.initials: C
authors.lastname: Even
authors.forename: G A
authors.initials: GA
authors.lastname: Palmer
authors.forename: P G
authors.initials: PG
authors.lastname: Plagemann
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D004220:Disulfides / Q000737:chemistry*; D007772:Lactate dehydrogenase-elevating virus / Q000378:metabolism / Q000472:pathogenicity*; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000502:physiology
chemical_list: D004220:Disulfides; D014759:Viral Envelope Proteins