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镁离子/钙离子 - ATP酶活性在从大鼠大脑皮层分离出的突触小泡中并未富集。

Mg2+/Ca(2+)-ATPase activity is not enriched in synaptic vesicles isolated from rat cerebral cortex.

作者信息

Rodríguez de Lores Arnaiz G, Pellegrino de Iraldi A

机构信息

Instituto de Biología Celular y Neurociencias Prof. Eduardo De Roberts Facultad de Medicina, Universidad de Buenos Aires, Argentina.

出版信息

Neurochem Res. 1997 Mar;22(3):293-6. doi: 10.1023/a:1022490822175.

Abstract

Neuronal ATPases comprise a wide variety of enzymes which are not uniformly distributed in different membrane preparations. Since purified vesicle fractions have Mg2+/Ca(2+)-ATPase, the purpose of the present study was to know whether such enzyme activities have a preferential concentration in a synaptic vesicle fraction in order to be used as markers for these organelles. Resorting to a procedure developed in this Institute, we fractionated the rat cerebral cortex by differential centrifugation following osmotic shock of a crude mitochondrial fraction and separated a purified synaptic vesicle fraction over discontinuous sucrose gradients. Mg2+/Ca(2+)-ATPase activities and ultrastructural studies of isolated fractions were carried out. It was observed that similar specific activities for Mg2+/Ca(2+)-ATPases were found in all fractions studied which contain synaptic vesicles and/or membranes. Although the present results confirm the presence of Mg2+ and Ca(2+)-ATPase activities in synaptic vesicles preparations, they do not favor the contention that Mg2+/Ca(2+)-ATPase is a good marker for synaptic vesicles.

摘要

神经元ATP酶包含多种酶,它们在不同的膜制剂中分布并不均匀。由于纯化的囊泡组分含有Mg2+/Ca(2+)-ATP酶,本研究的目的是了解这种酶活性是否在突触囊泡组分中具有优先浓度,以便用作这些细胞器的标记物。采用本研究所开发的方法,我们对大鼠大脑皮层进行分级分离,先对粗线粒体组分进行渗透压休克处理,然后通过差速离心法进行分离,并在不连续蔗糖梯度上分离出纯化的突触囊泡组分。对分离出的组分进行了Mg2+/Ca(2+)-ATP酶活性和超微结构研究。结果发现,在所研究的所有含有突触囊泡和/或膜的组分中,Mg2+/Ca(2+)-ATP酶的比活性相似。虽然目前的结果证实了突触囊泡制剂中存在Mg2+和Ca(2+)-ATP酶活性,但并不支持Mg2+/Ca(2+)-ATP酶是突触囊泡良好标记物的观点。

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