Integration of flux measurements and pharmacological controls to optimize stable isotope-resolved metabolomics workflows and interpretation.

ABSTRACT: Stable isotope-resolved metabolomics (SIRM) provides information regarding the relative activity of numerous metabolic pathways and the contribution of nutrients to specific metabolite pools; however, SIRM experiments can be difficult to execute, and data interpretation is challenging. Furthermore, standardization of analytical procedures and workflows remain significant obstacles for widespread reproducibility. Here, we demonstrate the workflow of a typical SIRM experiment and suggest experimental controls and measures of cross-validation that improve data interpretation. Inhibitors of glycolysis and oxidative phosphorylation as well as mitochondrial uncouplers serve as pharmacological controls, which help define metabolic flux configurations that occur under well-controlled metabolic sta

SUBMITTER: Lorkiewicz PK 

PROVIDER: S-EPMC6757038 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature