Enzymes are critical for life. Understanding how the atoms in enzymes move during the chemical reactions they catalyze is integral for the design of enzymes that would catalyze novel reactions and would lead to advances in the fields of biotechnology and drug design. This research aims to impact these fields through the characterization of enzyme motions and their importance to catalytic function. In addition to the direct impact on differing scientific areas, this project will result in the training of postdoctoral scientists in state-of-the-art biophysical techniques to enable their transition to independent researchers. Moreover, this project will recruit and train undergraduates, from underrepresented groups, in modern biophysical research.
The mechanism by which the catalytic activity of enzymes is regulated often involves the relay of structural or dynamical perturbations in the enzyme between a distant (allosteric) site and the catalytic (active) site. This project will examine the allosteric regulation of the human enzyme, protein tyrosine phosphatase 1B (PTP1B) using solution nuclear magnetic resonance (NMR) and computational methods coupled with biochemical experiments. These methods will enable examination of the kinetics, thermodynamics, and mechanism of motions in PTP1B. The goals of this project will be to understand how allosteric changes can be relayed over long molecular-scale distances and influence chemical reactivity. These goals will be achieved by characterizing how the binding of small, organic molecules at an allosteric site changes the structure and atomic flexibility of amino acids at the active site. In addition, studies will be performed to examine the natural allosteric site in PTP1B that is utilized in vivo to regulate its catalytic activity. This combination of experiments will provide new insight into PTP1B function and into general allosteric models applicable to numerous other important enzymes.
