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及蛋白质熔球态的方案。

Bose Himangshu S, Bose Mahuya, Whittal Randy M

Laboratory of Biochemistry and Cell Biology, Department of Biomedical Sciences, Mercer University School of Medicine, Savannah, GA 31404, USA; Anderson Cancer Institute, Memorial University Medical Center, Savannah, GA 31404, USA.

Center for Health Regeneration and Biotechnology and Department of Physiology and Functional Genomics, University of Florida, Gainesville, FL, USA.

STAR Protoc. 2025 Jul 25;6(3):103971. doi: 10.1016/j.xpro.2025.103971.

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, Bose et al., Pawlak et al., and Prasad et al..

类固醇激素对于所有哺乳动物在碳水化合物代谢、应激管理和有性生殖方面的生存至关重要。在此，我们展示了一种通过在类固醇生成和非类固醇生成细胞系统中测量孕烯醇酮或孕酮合成来评估线粒体胆固醇转运和蛋白质熔球状态的方案。我们描述了细胞培养、转染以及测量非类固醇生成细胞类固醇生成活性的步骤。然后详细说明了代谢转化的程序。有关本方案使用和执行的完整详细信息，请参考博斯、博斯等人、帕夫拉克等人以及普拉萨德等人的研究。