The price of flexibility – a case study on septanoses as pyranose mimetics

Abstract

Seven-membered ring mimetics of mannose were studied as ligands for the mannose-specific bacterial lectin FimH, which plays an essential role in the first step of urinary tract infections (UTI). A competitive binding assay and isothermal titration calorimetry (ITC) experiments indicated an approximately ten-fold lower affinity for the seven-membered ring mannose mimetic 2-<i>O-n</i>-heptyl-1,6-anhydro-d-<i>glycero</i>-d-galactitol (<b>7</b>) compared to <i>n</i>-heptyl α-d-mannopyranoside (<b>2</b>), resulting exclusively from a loss of conformational entropy. Investigations by solution NMR, X-ray crystallography, and molecular modeling revealed that <b>7</b> establishes a superimposable H-bond network compared to mannoside <b>2</b>, but at the price of a high entropic penalty due to the loss of its pronounced conformational flexibility. These results underscore the importance of having access to the complete thermodynamic profile of a molecular interaction to "rescue" ligands from entropic penalties with an otherwise perfect fit to the protein binding site.