Probing the binding mechanisms of ?-tocopherol to trypsin and pepsin using isothermal titration calorimetry, spectroscopic, and molecular modeling methods.

ABSTRACT: ?-Tocopherol is a required nutrient for a variety of biological functions. In this study, the binding of ?-tocopherol to trypsin and pepsin was investigated using isothermal titration calorimetry (ITC), steady-state and time-resolved fluorescence measurements, circular dichroism (CD) spectroscopy, and molecular modeling methods. Thermodynamic investigations reveal that ?-tocopherol binds to trypsin/pepsin is synergistically driven by enthalpy and entropy. The fluorescence experimental results indicate that ?-tocopherol can quench the fluorescence of trypsin/pepsin through a static quenching mechanism. The binding ability of ?-tocopherol with trypsin/pepsin is in the intermediate range, and one molecule of ?-tocopherol combines with one molecule of trypsin/pepsin. As shown by circular dichroism (CD) spectroscopy, ?-tocopherol may induce conformational changes of trypsin/pepsin. Molecular modeling displays the specific binding site and gives information about binding forces and ?-tocopherol-tryptophan (Trp)/tyrosine (Tyr) distances. In addition, the inhibition rate of ?-tocopherol on trypsin and pepsin was studied. The study provides a basic data set for clarifying the binding mechanisms of ?-tocopherol with trypsin and pepsin and is helpful for understanding its biological activity in vivo.

SUBMITTER: Li X 

PROVIDER: S-EPMC4942422 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature