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牛脑磷脂酰肌醇转移蛋白。pH值、离子强度和脂质组成对转移活性的影响。

Bovine brain phosphatidylinositol transfer protein. Effects of pH, ionic strength and lipid composition on transfer activity.

作者信息

Yoshimura T, Helmkamp G M

出版信息

Biochim Biophys Acta. 1984 May 11;793(3):463-70. doi: 10.1016/0005-2760(84)90264-9.

Abstract

Phosphatidylinositol and phosphatidylcholine are transferred between bilayer membranes in the presence of a specific phosphatidylinositol transfer protein isolated from bovine brain. The effects of pH, ionic strength and lipid composition on the rate of transfer of these phospholipids between small unilamellar vesicles have been investigated. At low ionic strength, phosphatidylinositol transfer between vesicles prepared from phosphatidylcholine and 5 mol% phosphatidylinositol was maximal at about pH 5 and moderately dependent on hydrogen ion concentration in more alkaline regions. A similar dependence on pH was noted for phosphatidylcholine transfer between membranes containing phosphatidylcholine or mixtures of phosphatidylcholine and 5 mol% phosphatidylinositol, phosphatidic acid, phosphatidylglycerol, phosphatidylethanolamine or stearylamine. The rate of transfer between anionic vesicles was somewhat higher than that between neutral or cationic vesicles. At higher ionic strength the transfer reactions in neutral and alkaline regions were less sensitive to pH. Phospholipid transfers between vesicles containing 5 mol% of anionic lipid increased sharply as ionic strength decreased below 0.1. In contrast, phosphatidylcholine transfer between membranes which contained only zwitterionic phospholipids or 5 mol% stearylamine was unaffected by variations of ionic strength. Irrespective of the lipid composition of membranes, pH affected both the apparent Km and Vmax, while ionic strength generally affected the apparent Vmax. These results indicate a significant role of electrostatic interactions in the phospholipid transfer catalyzed by phosphatidylinositol transfer protein.

摘要

在存在从牛脑中分离出的特定磷脂酰肌醇转移蛋白的情况下,磷脂酰肌醇和磷脂酰胆碱会在双层膜之间转移。已经研究了pH、离子强度和脂质组成对这些磷脂在小单层囊泡之间转移速率的影响。在低离子强度下,由磷脂酰胆碱和5摩尔%磷脂酰肌醇制备的囊泡之间的磷脂酰肌醇转移在约pH 5时最大,并且在更碱性区域中适度依赖于氢离子浓度。对于含有磷脂酰胆碱或磷脂酰胆碱与5摩尔%磷脂酰肌醇、磷脂酸、磷脂酰甘油、磷脂酰乙醇胺或硬脂胺混合物的膜之间的磷脂酰胆碱转移,也观察到了类似的对pH的依赖性。阴离子囊泡之间的转移速率略高于中性或阳离子囊泡之间的转移速率。在较高离子强度下,中性和碱性区域中的转移反应对pH的敏感性较低。当离子强度降至0.1以下时,含有5摩尔%阴离子脂质的囊泡之间的磷脂转移急剧增加。相比之下,仅含有两性离子磷脂或5摩尔%硬脂胺的膜之间的磷脂酰胆碱转移不受离子强度变化的影响。无论膜的脂质组成如何,pH都会影响表观Km和Vmax,而离子强度通常会影响表观Vmax。这些结果表明静电相互作用在磷脂酰肌醇转移蛋白催化的磷脂转移中起重要作用。

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