人类血小板中存在多种形式的鞘氨醇激酶的证据。
Evidence for the presence of multiple forms of Sph kinase in human platelets.
作者信息
Banno Y, Kato M, Hara A, Nozawa Y
机构信息
Department of Biochemistry, Gifu University School of Medicine, Tsukasamachi-40, Gifu 500-8705, Japan.
出版信息
Biochem J. 1998 Oct 15;335 ( Pt 2)(Pt 2):301-4. doi: 10.1042/bj3350301.
Abstract
The intracellular distribution of sphingosine (Sph) kinase activity was examined in human platelets. A large proportion (72%) of the total activity was found to be associated with the membrane fraction, and the membrane-associated fraction had higher specific activity compared with the cytosolic enzyme. Most of the membrane-associated activity could be extracted with 1 M NaCl. The cytosolic activity was unstable upon heat treatment, with 80% of the activity being lost during incubation at 45 degreesC for 1 h, whereas the NaCl-extractable fraction was stable under the same conditions. When subjected to Mono Q column chromatography, the cytosolic fraction produced two activity peaks and the NaCl-extractable fraction gave a single peak. These three Sph kinase activities showed different responses to stimulation by beta-octylglucoside and inhibition by N, N-dimethylsphingosine and l-threo-dihydrosphingosine, suggesting the presence of multiple enzyme forms in human platelets.
摘要
对人血小板中鞘氨醇（Sph）激酶活性的细胞内分布进行了检测。发现总活性的很大一部分（72%）与膜部分相关，且与胞质酶相比，膜相关部分具有更高的比活性。大部分膜相关活性可用1 M NaCl提取。胞质活性在热处理时不稳定，在45℃孵育1小时期间80%的活性丧失，而NaCl可提取部分在相同条件下是稳定的。当进行Mono Q柱层析时，胞质部分产生两个活性峰，而NaCl可提取部分给出一个单峰。这三种Sph激酶活性对β - 辛基葡糖苷的刺激以及对N，N - 二甲基鞘氨醇和l - 苏式 - 二氢鞘氨醇的抑制表现出不同的反应，提示人血小板中存在多种酶形式。
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