ARACHIDONOYL DIACYLGLYCEROL KINASE

花生四烯酰二酰甘油激酶

• 批准号: 6219613
• 负责人: John A. Glomset
• 金额: $ 5.72万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6219613
• 关键词: animal tissue; enzymes; human tissue; lipids; nervous system

We previously purified and characterized an arachidonoyl diacylglycerol kinase and showed that it is present in the testis, brain, and retina of baboons. The enzyme is one of at least ten diacylglycerol kinases that are present in mammalian tissues. We have now partially purified and characterized an additional diacylglycerol kinase, which is coexpressed with the arachidonoyl diacylglycerol kinase. The diacylglycerol kinase is soluble, but can bind to unilamellar lipid vesicles in vitro. Two types of binding sites seem to be involved, one that interacts with vesicle-associated acidic lipids and one that interacts with vesicle-associated diacylglycerol. Both types of binding sites must be engaged for effective enzyme-vesicle interaction. Under these conditions and when both Mg2+ and ATP are present in the medium, enzyme-vesicle interaction is optimal and correlates with catalysis. Even so, linear reaction rates are a complex function of what others have termed enzyme "scooting " and "hopping" phenomena. The enzyme apparently binds to the surface of a vesicle, catalyzes a few reactions, then dissociates from that surface, binds to a new vesicle, and begins a new round of catalysis. FUNDING NIH grant RR00166 and the Howard Hughes Medical Institute.

我们之前纯化并表征了花生四烯酰 二酰甘油激酶并表明它存在于睾丸中， 狒狒的大脑和视网膜。 该酶是至少十种酶之一 存在于哺乳动物组织中的二酰基甘油激酶。 我们有 现在部分纯化并表征了额外的二酰基甘油 激酶，与花生四烯酸二酰甘油共表达 激酶。 二酰甘油激酶是可溶的，但可以结合 体外单层脂质囊泡。 似乎有两种类型的结合位点 参与其中，与囊泡相关的酸性相互作用 脂质和一种与囊泡相关的二酰基甘油相互作用的脂质。 两种类型的结合位点都必须参与才能有效 酶-囊泡相互作用。 在这些条件下，当 Mg2+ 和 ATP 存在于培养基中，酶-囊泡相互作用是 最佳且与催化相关。 即便如此，线性反应速率 是其他人所说的酶“滑行”的复杂功能 和“跳跃”现象。 该酶显然与表面结合 囊泡，催化一些反应，然后从中解离 表面，与新的囊泡结合，并开始新一轮的催化。 资助 NIH 拨款 RR00166 和霍华德休斯医学研究所。