Isolation of the structural proteins of western equine encephalitis virus by isoelectric focusing.
Western equine encephalitis virus was disrupted with Triton X-100 and subjected to isoelectric focusing in a sucrose or urea gradient. The two envelope proteins, E1 and E2 were not well separated in a sucrose gradient, while the E1 and E2 proteins were distinguished as two major peaks which focused in a urea gradient at about pH 7.5 and 10, respectively. Isolated E1 protein refocused at pH 6.5 in a sucrose gradient isoelectric focusing column. When Western equine encephalitis virus was treated with Triton X-100 in 0.01 M phosphate buffer (pH6), hemagglutinating E1 protein was solubilized, which isoelectrofocused at pH 6.5. Purified nucleocapsids focused at pH 4 in a sucrose gradient on an isoelectric focusing column. After ribonuclease treatment of the purified nucleocapsid more than 95 per cent of the viral RNA was acid-soluble, and hte nucleocapsid protein isoelectrofocused at about pH 4.
D004667:Encephalitis Virus, Western Equine / Q000032:analysis* / Q000187:drug effects; D006389:Hemagglutinins, Viral / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods; D011092:Polyethylene Glycols / Q000494:pharmacology; D012367:RNA, Viral / Q000302:isolation & purification; D014764:Viral Proteins / Q000302:isolation & purification*
D006389:Hemagglutinins, Viral; D012367:RNA, Viral; D014764:Viral Proteins; D011092:Polyethylene Glycols