Biochemical analysis of bovine viral diarrhea virus polypeptides and studies of strain variation
Intracellular viral-specific polypeptides from the National Animal Disease Laboratory (NADL) strain of bovine viral diarrhea virus were studied by biosynthesis labelling, radioimmunoprecipitation (RIP), hypertonic initiation block (HIB) and polyacrylamide gel electrophoresis (PAGE). Eighteen virus-specific proteins were identified; thirteen were glycosylated (gp170, p135, p130, gp118, gp82, p80, gp74, gp63, gp60, p59, gp53, gp50, gp45, gp42, p37, gp32, gp25 and p22). When glycosylation was inhibited by tunicamycin, five {sup 35}S-methionine labelled proteins displayed increased electrophoretic mobility (gp170 to p165, gp74 to p66, gp53 to p45, gp50 to p42 and gp25 to p20) and four could not be identified. Similar shifts in mobility were observed following in vitro deglycosylation with endoglycosidases H and F indicating that the nine glycoproteins contained N-linked simple or high mannose containing moieties. Biosynthetic labelling in the presence of the ionophore, monensin, or in vitro deglycosylation with the endoglycosidase, O-glycanase, had no effect, which is consistent with the absence of O-linked carbohydrates in BVDV-specific proteins. N-linked glycosylation of BVDV proteins is critical for infectivity, because the virus from cells treated with tunicamycin was devoid of infectivity, whereas the virus from monensin-treated cells was fully infective. Partitioning of p130, p59, gp53-50, and p37 into solutions of Triton X-114 tentatively identified these molecules as partially hydrophobic transmembrane proteins. Biosynthesis in the presence of {sup 3}H-myristate and {sup 3}H-palmitate did not result in specifically labelled viral proteins indicating predominantly noncovalent nature of putative interactions of these proteins with membranes. Partial proteolytic peptide mapping revealed similarities among gp170, p130 and p80 and between gp53 and gp50.
- Research Organization:
- Colorado State Univ., Fort Collins, CO (United States)
- OSTI ID:
- 5602834
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOASSAY
BIOLOGICAL VARIABILITY
CARBOXYLIC ACIDS
CATTLE
CHEMICAL ANALYSIS
DAYS LIVING RADIOISOTOPES
DIAGNOSTIC TECHNIQUES
DISEASES
DOMESTIC ANIMALS
DRUGS
ELECTROPHORESIS
EVEN-ODD NUCLEI
GENETIC VARIABILITY
HYDROGEN COMPOUNDS
IMMUNOASSAY
IMMUNOLOGY
INFECTIOUS DISEASES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
METHIONINE
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PEPTIDES
POLYPEPTIDES
PROTEINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNODETECTION
RADIOIMMUNOLOGY
RADIOISOTOPES
RUMINANTS
SULFUR 35
SULFUR ISOTOPES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES
VIRAL DISEASES