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.
D000217:Acyltransferases / Q000276:immunology / Q000302:isolation & purification*; D019881:Aminoacyltransferases; D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005779:Immunodiffusion; D007668:Kidney / Q000201:enzymology*; D011817:Rabbits; D013552:Swine
D000217:Acyltransferases; D019881:Aminoacyltransferases; C019727:arginyltransferase