Envelope glycoproteins of Rauscher murine leukemia virus: isolation and chemical characterization
作者:Hans Marquardt, Raymond V. Gilden, Stephen I. Oroszlan · 发表于:Biochemistry · 年份:1977 · DOI:10.1021/bi00623a024 · 被引用次数:56 · 研究领域:Glycosylation and Glycoproteins Research、Monoclonal and Polyclonal Antibodies Research、T-cell and Retrovirus Studies
The envelope glycoproteins (designated gp70 and gp45) of the Rauscher strain of murine leukemia virus were solubilized by osmotic shock and freeze-thawing in chaotropic solutions. The viral glycoproteins were then purified by phosphocellulose chromatography and gel permeation chromatography on Bio-Gel A-1.5m. Yields by this procedure were 6.2% for gp70 and 1.3% for gp45 on a protein input basis. The apparent molecular weights were respectively 67 500 and 47 500 with a polypeptide chain molecular weight of approximately 45 000 for both glycoproteins. Amino acid analysis showed a high degree of similarity for both components, with some differences subject to further evaluation. The total carbohydrate content was approximately 32% for gp70 and 6-9% for gp45. In keeping with the amino acid compositional similarity suggesting relationships, alanine was found to ba the amino-terminal amino acid of both glycoproteins, and cross-reactivity was demonstrated by immunologic tests. The data suggest that the chief difference between gp70 and gp45 lies in the carbohydrate content.