Semicarbazide-sensitive amine oxidase in pig heart
A semicarbazide-sensitive amine oxidase (SSAO) (E.C.188.8.131.52) has been purified from pig heart. Western blot analysis showed that the enzyme reacts with a polyclonal antibody raised against homogeneous crystalline pig plasma benzylamine oxidase (BAO). A subunit molecular mass of 97 KDa obtained by SDS electrophoresis is identical to the plasma enzyme. The purification procedure consisted of sequential DEAE cellulose, octyl-Sepharose, Con A-Sepharose and hydroxyapatite columns. Two peaks of activity were obtained on octyl-Sepharose which were found to be kinetically and immunologically indistinguishable. The specific activity of the purified enzyme was 0.045 (imol/min/mg of protein at 37°C and the Km for benzylamine was estimated to be 63 μM. The enzyme was inhibited by carbonyl reagents such as semicarbazide but was insensitive to the effect of pargyline.
KeywordsAmmonium Sulphate DEAE Cellulose Amine Oxidase SSAO Activity 50mM Phosphate Buffer
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