Properties of Feline Leukemia Virus I. Chromatographic Separation and Analysis of the Polypeptides

Abstract

Rickard's strain of feline leukemia virus (FeLV) contains two large glycoproteins and five smaller polypeptides of molecular weights 100,000 (gp ≥ 100), 70,000 (gp70), 30,000 (p30), 21,000 (p21), 15,000 (p15), 11,200 (p11), and 10,000 (p10) when chromatographed on 6% agarose in the presence of 6 M guanidine hydrochloride (GuHCl). P21 is a minor component which was not previously described for mammalian leukemia-sarcoma viruses and may be analogous to the seventh protein found in avian viruses. Analysis on 4% agarose and by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that gp ≥ 100 is actually ≥ 200,000 daltons and dissociates to polypeptides of approximately 100,000 to 115,000 daltons, whereas gp70 can be resolved into six stained bands ranging from 40,000 to 80,000 daltons despite being rechromatographed as a single symmetrical peak on 6% agarose. Rechromatography on 8% agarose was found to be more effective than on 6% agarose or sodium dodecyl sulfate polyacrylamide gel electrophoresis for obtaining the five small polypeptides, especially p11 and p10, in a highly purified form suitable for further analysis and for obtaining more precise estimates of their molecular weights, especially when done by co-chromatography with iodinated standard proteins markers. Rechromatographed p30, p21, p15, p11, and p10 had molecular weights of 27,000, 18,000, 15,000, 12,000, and 12,000 respectively, by co-electrophoresis with the marker proteins on sodium dodecyl sulfate polyacrylamide gel electrophoresis, clearly establishing that the latter two FeLV polypeptides comigrate to form one less band when compared to elution from agarose. The isoelectric points of p30 and p15 were 5.5 and 8.9, respectively, after renaturation from GuHCl and 5.6 and 8.7, respectively, when isolated from Tween-ether treated virus. Rechromatographed p30, p15, and p11, renatured by removing GuHCl with dialysis, reacted only with their homologous antisera in immunodiffusion analysis, indicating that they are immunologically unrelated. Also the interspecies gs-3 determinant associated with p30 could be regained by removal of GuHCl.