Mechanism of action of steroidogenic acute regulatory protein (StAR)

描述（由申请人提供）：类固醇生成的限速步骤是胆固醇从线粒体外膜（OMM）向线粒体内膜（IMM）的移动。类固醇生成急性调节蛋白（星星）通过一种未知的机制促进胆固醇从OMM向IMM的移动，从而调节底物向P450 SCC的流动以进行类固醇生成。星星属于称为START（星星相关脂质转运结构域）的蛋白质家族，其中星星是主要成员。星星突变会导致一种潜在的致命疾病，称为先天性肾上腺增生，其中所有类固醇激素的合成都受到损害，胎儿在出生后不久就会死亡。星星资产举措的活动与其在监察员办公室的驻留时间成正比。我们的初步结果表明，i）星星的输入机制是独特的，ii）星星暂停序列在星星折叠和活性中具有独特的作用，iii）星星在一小时内转运超过100个胆固醇分子，iv）星星通过几种蛋白质的复合物输入，v）星星以两步过程输入。为了了解星星加工和输入线粒体的机制，我们提出了三个具体目标。目的1：研究新合成的星星的活性机制。这对于理解星星如何以如此短的半衰期发挥作用以及它如何在没有N-末端序列的情况下到达OMM至关重要。目的二是确定StAR在线粒体中的作用位点，以了解星星通过OMM驻留蛋白VDAC 1和VDAC 2（电压依赖性阴离子通道）的转运机制。在目标3中，我们提出通过OMM相关的脂质膜来确定星星的解折叠机制，并揭示为什么特定的START结构域比其他结构域更活跃。实现这些目标将有助于我们扩大对星星与OMM相互作用机制的了解，以及扩大对OMM作用的了解，OMM有助于星星展开，从而动员胆固醇转运。完成这些目标将有助于我们了解细胞脂质代谢的广泛方面，以及广泛的出生缺陷，胚胎发育障碍和遗传疾病。

DESCRIPTION (provided by applicant): The rate-limiting step in steroidogenesis is the movement of cholesterol from the outer mitochondrial membrane (OMM) to inner mitochondrial membrane (IMM). The steroidogenic acute regulatory protein (StAR) by an unknown mechanism facilitates the movement of cholesterol from the OMM to IMM, thus regulating the flow of substrate to P450scc for steroidogenesis. StAR belongs to a family of proteins called START (StAR related lipid transport domain), where StAR is the primary member. StAR mutations cause a potentially lethal disease called congenital adrenal hyperplasia, in which the synthesis of all steroid hormones is impaired, and the fetuses die shortly after birth. StAR activity is directly proportional to its residency time at the OMM. Our preliminary results show that i) the import mechanism of StAR is unique, ii) the StAR pause sequence has a distinct role in StAR folding and activity, iii) StAR transports more than 100 molecules of cholesterol in one hour, iv) StAR is imported through a complex of several proteins, and v) StAR is imported in a two step process. To understand the mechanism of StAR processing and import into mitochondria, we propose three specific aims. Aim 1 is to determine the mechanism of activity of newly synthesized StAR. This is critical in understanding how StAR functions with such a short half- life, and how it can reach the OMM with out the N-terminal sequence. In aim 2, we proposed to determine the mitochondrial site of StAR action, in order to understand the translocation mechanism of StAR by OMM resident proteins VDAC 1 and 2 (Voltage Dependent Anion Channel). In aim 3, we proposed to determine the unfolding mechanism of StAR by the OMM associated lipid membrane, and to uncover why a specific START-domain is more active than the other. Fulfilling these aims will, I) help us to expand our knowledge about the mechanism of interaction of StAR with the OMM and II) expand our understanding about the role of OMM that help in StAR unfolding and thus mobilization of cholesterol transport. Completing these aims will help us in the understanding of broad aspects of cellular lipid metabolism and also to a broad array of birth defects, disorders of embryogenesis and genetic diseases.