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猪心脏中的氨基脲敏感胺氧化酶

Semicarbazide-sensitive amine oxidase in pig heart.

作者信息

Dowling T G, Cambi S, Buffoni F

机构信息

Department of Pharmacology, University of Florence, Italy.

出版信息

J Neural Transm Suppl. 1998;52:265-70. doi: 10.1007/978-3-7091-6499-0_26.

Abstract

A semicarbazide-sensitive amine oxidase (SSAO) (E.C.1.4.3.6) 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 mumol/min/mg of protein at 37 degrees C and the Km for benzylamine was estimated to be 63 microM. The enzyme was inhibited by carbonyl reagents such as semicarbazide but was insensitive to the effect of pargyline.

摘要

已从猪心中纯化出一种氨基脲敏感胺氧化酶(SSAO)(E.C.1.4.3.6)。蛋白质免疫印迹分析表明,该酶与针对纯化的猪血浆苄胺氧化酶(BAO)产生的多克隆抗体发生反应。通过SDS电泳获得的97 kDa亚基分子量与血浆酶相同。纯化过程包括依次使用DEAE纤维素柱、辛基琼脂糖柱、伴刀豆球蛋白A琼脂糖柱和羟基磷灰石柱。在辛基琼脂糖柱上获得了两个活性峰,发现它们在动力学和免疫学上无法区分。纯化酶在37℃时的比活性为0.045 μmol/分钟/毫克蛋白质,苄胺的Km估计为63 μM。该酶被氨基脲等羰基试剂抑制,但对炔丙基胺的作用不敏感。

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