Complete purification of arginyl-tRNA:protein arginyltransferase from hog kidney and production of its antibody.

Arginyltransferase was purified 60,000-fold from the 105,000g supernatant of hog kidney by fractionation with ammonium sulfate, precipitation at pH 5.2, and chromatographies on DEAE-cellulose and hydroxyapatite, as well as affinity chromatographies on heparin-Sepharose and angiotensin II-Sepharose. Purified transferase had a specific activity of 7.6 mumol/min/mg, which was 66 times higher than that attained thus far (R. L. Soffer, 1970, J. Biol. Chem. 245, 731-737). Electrophoresis of purified transferase on polyacrylamide gel showed a single band which corresponded to the region of activity in homogenized slices of a duplicate gel. The homogeneity was also observed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, where only a single band of Mr 35,000 was observed, even after staining by the silver method (B. R. Oakley, D. R. Kirsch, and N. R. Morris, 1980, Anal. Biochem. 105, 361-363). Antibody to the purified transferase was prepared in a rabbit. The antibody inhibited arginyltransferase activity, and showed a single precipitin line by double immunodiffusion with crude transferase preparation.