评估类固醇生成和非类固醇生成系统中线粒体胆固醇转运和蛋白质熔融球状态的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-07-25 DOI:10.1016/j.xpro.2025.103971

Himangshu S Bose, Mahuya Bose, Randy M Whittal

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引用次数: 0

摘要

类固醇激素对所有哺乳动物的生存至关重要，用于碳水化合物代谢、压力管理和有性繁殖。在这里，我们提出了一种通过测量孕烯醇酮或孕酮合成在甾体和非甾体细胞系统中评估线粒体胆固醇转运和蛋白质熔融球状态的方案。我们描述了细胞培养、转染和测量非甾体生成细胞的甾体生成活性的步骤。然后详细介绍代谢转化的程序。有关本协议使用和执行的完整细节，请参阅Bose，1 Bose等人，2 Pawlak等人，3和Prasad等人。

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Protocol for assessing mitochondrial cholesterol transport and protein molten globule state in steroidogenic and nonsteroidogenic systems.

Steroid hormones are essential for the survival of all mammals for carbohydrate metabolism, stress management, and sexual reproduction. Here, we present a protocol for assessing mitochondrial cholesterol transport and protein molten globule state via measurement of pregnenolone or progesterone synthesis in steroidogenic and nonsteroidogenic cellular systems. We describe steps for cell culture, transfection, and measurement of steroidogenic activity from nonsteroidogenic cells. We then detail procedures for metabolic conversion. For complete details on the use and execution of this protocol, please refer to Bose,¹ Bose et al.,² Pawlak et al.,³ and Prasad et al.⁴.

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