Trends in thermostability provide information on the nature of substrate, inhibitor, and lipid interactions with mitochondrial carriers.

ABSTRACT: Mitochondrial carriers, including uncoupling proteins, are unstable in detergents, which hampers structural and mechanistic studies. To investigate carrier stability, we have purified ligand-free carriers and assessed their stability with a fluorescence-based thermostability assay that monitors protein unfolding with a thiol-reactive dye. We find that mitochondrial carriers from both mesophilic and thermophilic organisms exhibit poor stability in mild detergents, indicating that instability is inherent to the protein family. Trends in the thermostability of yeast ADP/ATP carrier AAC2 and ovine uncoupling protein UCP1 allow optimal conditions for stability in detergents to be established but also provide mechanistic insights into the interactions of lipids, substrates, and inhibitors with t

SUBMITTER: Crichton PG 

PROVIDER: S-EPMC4375477 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature