Active site voltage clamp fluorometry of the sodium glucose cotransporter hSGLT1.

ABSTRACT: In the human sodium glucose cotransporter (hSGLT1) cycle, the protein undergoes conformational changes where the sugar-binding site alternatively faces the external and internal surfaces. Functional site-directed fluorometry was used to probe the conformational changes at the sugar-binding site. Residues (Y290, T287, H83, and N78) were mutated to cysteines. The mutants were expressed in Xenopus laevis oocytes and tagged with environmentally sensitive fluorescent rhodamines [e.g., tetramethylrhodamine (TMR)-thiols]. The fluorescence intensity was recorded as the mutants were driven into different conformations using voltage jumps. Sugar binding and transport by the fluorophore-tagged mutants were blocked, but Na⁺ binding and the voltage-dependent conformational transitions

SUBMITTER: Gorraitz E 

PROVIDER: S-EPMC5699082 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature