Screening of Saccharomyces cerevisiae metabolite transporters by ¹³C isotope substrate labeling.

ABSTRACT: The transportome of Saccharomyces cerevisiae comprises approximately 340 membrane-bound proteins, of which very few are well-characterized. Elucidating transporter proteins' function is essential not only for understanding central cellular processes in metabolite exchange with the external milieu but also for optimizing the production of value-added compounds in microbial cell factories. Here, we describe the application of ¹³C-labeled stable isotopes and detection by targeted LC-MS/MS as a screening tool for identifying Saccharomyces cerevisiae metabolite transporters. We compare the transport assay's sensitivity, reproducibility, and accuracy in yeast transporter mutant cell lines and Xenopus oocytes. As proof of principle, we analyzed the transport profil

SUBMITTER: Stanchev LD 

PROVIDER: S-EPMC10729909 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature