| Abstract: | A ribonuclease inhibitor from bovine brain has been purified 27000-fold by affinity chromatography on Sepharose-RNase A with an overall yield of 46% (ca. 0.4 mg/kg tissue). The purified inhibitor gives a single band by SDS-gel electrophoresis. By gel filtration the molecular weight is ca. 50 000; the molecular weight of the complex with bovine pancreatic RNase A is ca. 62 000, indicative of 1:1 binding on a molar basis. The inhibition of the action of RNase A on yeast RNA by the inhibitor is noncompetitive with a Kiof 7 times 10-10 M. The protein is very similar in its properties, including amino acid composition, to the inhibitor previously isolated from human placenta. The amount of inhibitor per g of protein in bovine brain is about one-seventh of the value for human placenta. No difference was found in the distribution of inhibitor between white and gray matter; one-tenth of the inhibitor present is bound to a brain ribonuclease which is released in active form after reaction with p-hydroxymercuribenzoate. Essentially no free neutral ribonuclease activity could be detected in brain homogenates in the absence of p-hydroxymercuribenzoate. |
