Measuring CPT-1-mediated respiration in permeabilized cells and isolated mitochondria.

ABSTRACT: 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).

SUBMITTER: Yang K 

PROVIDER: S-EPMC8348308 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature