Purification of viral proteins from avian sarcoma virus QV2

• Published in: Journal of biochemistry (Tokyo) 1979-01, Vol.86 (4), p.929-942
• Main Authors: UENO, Akemichi, KOHNO, Michiaki, ISHIHAMA, Akira, TOYOSHIMA, Kumao
• Format: Article
• Language: English
• Subjects: DNA-Directed DNA Polymerase - isolation & purification; Immunodiffusion; Membrane Proteins - isolation & purification; Molecular Weight; Retroviridae - analysis; Retroviridae - enzymology; Viral Proteins - isolation & purification
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/oxfordjournals.jbchem.a132625

A procedure was established whereby most of the major viral proteins were isolated to apparent homogeneity in biologically and immunologically active forms from a single batch of avian sarcoma virus QV2. For the initial step of purification, gently disrupted virions were fractionated by CsCl centrifugation into envelope proteins, RNA-dependent DNA polymerase, and viral core proteins. Further purification of envelope glycoproteins and DNA polymerase was performed by affinity chromatography on agarose columns cross-linked with plant lectins and poly(C), respectively. On the other hand, core proteins were fractionated by a combination of gel filtration and ion-exchange column chromatography into components p27, pl9, and p15. The core protein p15 thus isolated retained proteolytic activity even after storage for 6 months. The present study also demonstrated that QV2 p19 is structurally altered from the corresponding protein of avian myeloblastosis virus (AMV), a reference avian leukosis-sarcoma virus having a well-characterized polypeptide composition.