Oversized galactosides as a probe for conformational dynamics in LacY.

ABSTRACT: Binding kinetics of α-galactopyranoside homologs with fluorescent aglycones of different sizes and shapes were determined with the lactose permease (LacY) of Escherichia coli by FRET from Trp151 in the binding site of LacY to the fluorophores. Fast binding was observed with LacY stabilized in an outward-open conformation (k_on = 4-20 μM^-1·s^-1), indicating unobstructed access to the binding site even for ligands that are much larger than lactose. Dissociation rate constants (k_off) increase with the size of the aglycone so that K_d values also increase but remain in the micromolar range for each homolog. Phe27 (helix I) forms an apparent constriction in the pathway for sugar by protruding into the periplasmic ca

SUBMITTER: Smirnova I 

PROVIDER: S-EPMC5910868 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature