豚鼠脑中三磷酸肌醇磷酸单酯酶的亚细胞分布
The subcellular distribution of triphosphoinositide phosphomonoesterase in guinea-pig brain.
作者信息
Sheltawy A, Brammer M, Borrill D
出版信息
Biochem J. 1972 Jul;128(3):579-86. doi: 10.1042/bj1280579.
Abstract
- Some properties of the triphosphoinositide phosphomonoesterase from the homogenates of guinea-pig brain were studied. The enzyme has an optimum pH range 6.7-7.3, is stimulated with KCl at a concentration of 0.1m, and under these conditions has K(m)1.43x10(-4)m. 2. A factor from the ;pH5 supernatant' of guinea-pig brain stimulates the enzyme activity over and above the stimulation produced by KCl. Subcellular fractions of guinea-pig brain varied in their response to the ;pH5 supernatant'. Maximum stimulation was observed with the P(1) fraction, containing myelin and nuclei. 3. An assay system for the enzyme was developed that contained optimum concentrations of both KCl and the ;pH5 supernatant'. Acid phosphatases were inhibited by NaF, but, in contrast with previous work, no EDTA was added to the assay system to inhibit the alkaline phosphatases. This reagent inhibited the triphosphoinositide phosphomonoesterase. It was estimated that the remaining fraction of non-specific phosphatases can account for only 14% of the observed triphosphoinositide phosphomonoesterase activity. 4. Subcellular fractions of guinea-pig brain were characterized by electron microscopy and subcellular markers. The triphosphoinositide phosphomonoesterase exhibited a distribution between the fractions similar to that of 5'-nucleotidase, but different from that of alkaline phosphatase.
摘要
- 对豚鼠脑匀浆中三磷酸肌醇磷酸单酯酶的一些性质进行了研究。该酶的最适pH范围为6.7 - 7.3，在0.1m浓度的KCl作用下受到刺激，在此条件下其米氏常数K(m)为1.43×10(-4)m。2. 来自豚鼠脑“pH5上清液”的一种因子对该酶活性的刺激作用超过了KCl所产生的刺激。豚鼠脑的亚细胞组分对“pH5上清液”的反应各不相同。在含有髓磷脂和细胞核的P(1)组分中观察到最大刺激作用。3. 开发了一种该酶的测定系统，其中含有最佳浓度的KCl和“pH5上清液”。酸性磷酸酶被NaF抑制，但与先前的工作不同，测定系统中未添加EDTA来抑制碱性磷酸酶。这种试剂抑制了三磷酸肌醇磷酸单酯酶。据估计，非特异性磷酸酶的剩余部分仅占所观察到的三磷酸肌醇磷酸单酯酶活性的14%。4. 通过电子显微镜和亚细胞标志物对豚鼠脑的亚细胞组分进行了表征。三磷酸肌醇磷酸单酯酶在各组分之间的分布与5'-核苷酸酶相似，但与碱性磷酸酶不同。
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