测量透化细胞和分离的线粒体中的 CPT-1 介导的呼吸作用。
Measuring CPT-1-mediated respiration in permeabilized cells and isolated mitochondria.
机构信息
Department of Molecular and Medical Pharmacology, UCLA David Geffen School of Medicine, Los Angeles, CA 90095, USA.
Department of Medicine, UCLA David Geffen School of Medicine, Los Angeles, CA 90095, USA.
出版信息
STAR Protoc. 2021 Jul 31;2(3):100687. doi: 10.1016/j.xpro.2021.100687. eCollection 2021 Sep 17.
Carnitine palmitoyltransferase-1 (CPT-1) is a rate-controlling enzyme for long-chain fatty acid oxidation. This manuscript provides protocols for measuring CPT-1-mediated respiration in permeabilized, adherent cell monolayers and mitochondria freshly isolated from tissue, along with examples to assess the potency and specificity of interventions targeting CPT-1. Strengths of the approach include ease, speed, and breadth of analysis, whereas drawbacks include loss of physiological regulation in reductionist systems and indirect assessment of CPT-1 enzymatic activity. For complete details on the use and execution of this protocol, please refer to Divakaruni et al. (2018).
肉碱棕榈酰基转移酶-1（CPT-1）是长链脂肪酸氧化的限速酶。本手稿提供了在透化的贴壁细胞单层和组织中新鲜分离的线粒体中测量 CPT-1 介导的呼吸的方案，以及评估针对 CPT-1 的干预措施的效力和特异性的示例。该方法的优点包括易于操作、速度快且分析范围广，缺点包括在简化系统中丧失生理调节以及对 CPT-1 酶活性的间接评估。有关该方案使用和执行的完整详细信息，请参阅 Divakaruni 等人。（2018 年）。
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