Mach O, Sovová V, Cerná H
Folia Biol (Praha). 1978;24(4):233-41.
The isoelectric focusing technique in the pH gradients was used for a preparative isolation of proteins from Rous sarcoma virus and avian myeloblastosis virus. The purified major gs protein, p27 (pI = 9.1) and the gP86 (pI = 5.3) were obtained after disruption of virus with 1% non-ionogenic detergent in the presence of 6M urea. The p10, p15, and p19 were present in the same range of pH (pI = 6.8). A strongly basic protein, immunologically active, presumably the p12, was found in the alkaline region of the pH gradient 10.8. These proteins fully retained their immunological activity. On the other hand, in the acidic region of the pH gradient between pH 4 and 5, strong precipitates were regularly found. These precipitates were complexes which were formed by interaction of the acid components of ampholines with the viral proteins during isoelectric focusing. Almost all viral proteins were present, differing only in quantity. The complexes were stabile in 1% non-inogenic detergent and 6M urea. They were dissociated with 1% SDS and 5M urea, and had no immunological activity. The methods of virus disruption and possibilities of formation of the ampholine-viral protein complexes are discussed.
采用pH梯度等电聚焦技术从劳氏肉瘤病毒和禽成髓细胞瘤病毒中制备性分离蛋白质。在6M尿素存在下，用1%非离子型去污剂裂解病毒后，获得了纯化的主要gs蛋白p27（等电点=9.1）和gP86（等电点=5.3）。p10、p15和p19存在于相同的pH范围（等电点=6.8）。在pH梯度为10.8的碱性区域发现了一种具有免疫活性的强碱性蛋白，推测为p12。这些蛋白质完全保留了它们的免疫活性。另一方面，在pH 4至5的pH梯度酸性区域，经常发现强沉淀。这些沉淀是在等电聚焦过程中，两性电解质的酸性成分与病毒蛋白相互作用形成的复合物。几乎所有病毒蛋白都存在，只是数量不同。这些复合物在1%非离子型去污剂和6M尿素中稳定。它们用1% SDS和5M尿素解离，且无免疫活性。讨论了病毒裂解方法以及两性电解质-病毒蛋白复合物的形成可能性。
