PURIFICATION OF BOVINE PINEAL HYDROXYINDOLE O‐methylTRANSFERASE BY IMMUNOADSORPTION CHROMATOGRAPHY
PURIFICATION OF BOVINE PINEAL HYDROXYINDOLE O‐methylTRANSFERASE BY IMMUNOADSORPTION CHROMATOGRAPHY
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免疫吸附色谱法纯化牛松果体羟吲哚邻位甲基转移酶
DOI：
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发表时间：
1978
影响因子：
4.7
通讯作者：
Y. Takahashi
中科院分区：
文献类型：
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作者：
R. Kuwano;Y. Yoshida;Y. Takahashi
本文介绍了羟基吲哚O-甲基转移酶（HIOMT）的免疫吸附层析法。通过免疫球蛋白（IG）-琼脂糖凝胶亲和色谱法从牛松果体提取物中纯化HIOMT。总纯化约45倍;产率为84%。这种酶约占松果体中可溶性蛋白质的2.0%。该酶在Ouchterlony双扩散板和免疫电泳上呈现单一的沉淀线。超离心分析表明酶分子聚集体的存在，圆盘凝胶电泳显示一条主要蛋白带和几条次要蛋白带。然而，十二烷基硫酸钠（SDS）凝胶电泳显示亚基分子量为38，000的单一蛋白条带，表明牛松果体HIOMT是单一亚基的聚合酶。纯化酶的圆盘凝胶电泳图谱、pH、化学物质和底物的影响以及免疫学性质与粗酶相同。
A procedure is described for the use of immunoadsorption chromatography of hydroxyindole O‐methyltransferase (HIOMT). HIOMT was purified from bovine pineal extract by affinity chromatography on immunoglobulins (Ig)‐Sepharose. The overall purification was about 45‐fold; the yield was 84%. This enzyme constitutes about 2.0% of the soluble proteins in the pineal gland. The enzyme represented a single precipitin line on Ouchterlony double diffusion plate and immunoelectrophoresis. Ultracentrifugation analysis indicated the existence of molecular aggregates of enzyme and disc gel electrophoresis showed one main protein band and several minor bands. However sodium dodecyl sulphate (SDS) gel electrophoresis showed a single protein band with subunit molecular weight 38,000 demonstrating bovine pineal HIOMT to be polymer enzyme of a single subunit. The properties of the purified enzyme including disc gel electrophoretic pattern, the effect of pH, chemicals and substrates and immunological properties were identical with those of the crude enzyme.