title: Virion associated proteins of equine rhinitis B virus 1 (ERBV1): the non-structural protein 3C(pro) co-purifies with virions.
abstract: Equine rhinitis B virus (ERBV), genus Erbovirus, is most closely related to the Cardiovirus genus in the family Picornaviridae. The structural proteins (VP1-4) of erboviruses are not well described, but are predicted by sequence to be 35, 29, 26 and 7 kDa. Methods for the purification of cardioviruses (polyethylene glycol, trypsin treatment) were used to characterise the structural proteins of ERBV1. Only one of the virus proteins detected was an expected molecular mass, and this 26 kDa protein was identified as VP3 by N-terminal amino acid sequencing. N-terminal sequencing of the 56 and a 29 kDa protein identified sequences consistent with VP2 and VP1 respectively, despite these being 27 kDa larger and 6 kDa smaller than predicted. Virus purified without trypsin showed proteins more consistent with masses predicted for VP1, VP2 and VP3 at 35, 29 and 26 kDa respectively. These proteins were further identified with antibodies affinity purified to recombinant VP1, VP2, VP3 produced in E. coli. Interestingly, antibodies affinity purified to the non-structural protein 3C(pro), produced in insect cells, strongly detected a 27 kDa protein in western blots of virus purified with and without trypsin treatment, suggesting the non-structural 27 kDa 3C(pro) co-purifies with ERBV1 virions.
authors.affiliation: Centre for Equine Virology, School of Veterinary Science, The University of Melbourne, Victoria 3010, Australia.
authors.forename: Wesley D
authors.initials: WD
authors.lastname: Black
authors.forename: Carol A
authors.initials: CA
authors.lastname: Hartley
authors.forename: Nino P
authors.initials: NP
authors.lastname: Ficorilli
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Studdert
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000378:metabolism; D002522:Chlorocebus aethiops; D052698:Erbovirus / Q000235:genetics*; D008970:Molecular Weight; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000302:isolation & purification*; D014771:Virion / Q000235:genetics*
chemical_list: D000914:Antibodies, Viral; D017361:Viral Nonstructural Proteins