期刊
CELLULAR AND MOLECULAR LIFE SCIENCES
卷 65, 期 12, 页码 1948-1956出版社
SPRINGER BASEL AG
DOI: 10.1007/s00018-008-8206-z
关键词
orexin; hypocretin; phospholipase C; phospholipase D; phosphoinositides; phosphatidylcholine; diacylglycerol; live cell imaging
We investigated coupling of OX1 receptors to phospholipase activation and diacylglycerol generation in Chinese hamster ovary (CHO) cells using both biochemical and fluorescence real-time methods. The results indicate that at lowest orexin-A concentrations (highest potency), diacylglycerol generated results from phospholipase D activity. At 10-100-fold higher orexin-A concentrations, phospholipase C is activated, likely hydrolyzing phosphatidylinositol (PI) or phosphatidylinositol monophosphate (PIP) but not phosphatidylinositol bisphosphate (PIP2). At further 7-fold higher orexin-A concentrations, PIP2 is hydrolyzed, releasing both diacylglycerol and inositol-1,4,5-trisphosphate. Thus, OX1 orexin receptors connect to multiple phospholipase activities, apparently composed of at least one phospholipase D and two different phospholipase C activities. At low agonist concentrations, diacylglycerol and phosphatidic acid are the preferred products, and interestingly, it seems that even the primarily activated phospholipase C mainly works to increase diacylglycerol and not inositol-1,4,5-trisphosphate.
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