# Rapid protein dynamics and catalysis: modulation by laboratory evolution, designed mutation, and protein control of electric field environment

> **NIH NIH R01** · UNIVERSITY OF ARIZONA · 2020 · $297,920

## Abstract

The goal of the research program described in this application is to obtain a deeper understanding of how rapid
protein dynamics, so called promoting vibrations on a sub picosecond timescale, are employed and
incorporated into both natural and artificially designed enzymes. Over the years we have identified such
motions in a variety of enzymatically catalyzed reactions, and also found them missing in at least one. Our goal
for the long term is to decipher how nature has used this approach as an engineering principle and how it has
been incorporated as a part of the function. For example, many studies have now found that in hydride transfer
enzymes such as alcohol dehydrogenase, rapid motion of the donor to the acceptor facilitates the chemical
step by lowering the effective adiabatic barrier. We also study two seemingly dichotomous views of enzyme
function – the electrostatic view, and the dynamic view. It is entirely possible that the electric field milieu both
contributes strongly to catalysis, and in certain cases is modulated by the same types of motions that for
example control donor acceptor distance. Because our methods allow us to harvest ensembles of reactive
trajectories, exactly such mechanisms can be studied. Finally, there exist cases in which naturally occurring
enzymes exhibit simple and direct chemistry, while single mutations cause significant complexities in kinetic
analysis. It is simply not clear how this can come about. In order to pursue this research we propose to
implement the following three specific aims:
Aim 1: We will study one of the more successful attempts in synthetic enzymatic chemistry – the artificial
creation of retro-aldolases. We will ascertain whether theoretical design coupled to laboratory evolution of
these artificial protein catalysts caused change in the coupling of protein dynamics to reaction.
Aim 2: We will study how electric field varies in the active site of an enzyme as the reaction proceeds from
reactants to products through a transition state. Catechol-O-methyltransferase (COMT) is an enzyme in which
both protein dynamics and electrostatic preorganization have been stated emphatically by other groups to
produce the catalytic effect. In particular we will identify if there is a promoting vibration as part of the reaction
coordinate in this enzyme, and how it may help to control the field environment.
Aim 3: We will analyze reactions catalyzed by the poorly understood “ene-reductase” family. We will address
the importance of protein dynamics and the ability of a single point mutation to create multiple reaction
“configurations” with highly divergent kinetic behavior.
After decades of study, the deceptively simple question of how enzymes work is still a hotbed of debate. Via
such studies as here proposed we contribute to both basic knowledge and eventual practical control
application.

## Key facts

- **NIH application ID:** 9839628
- **Project number:** 5R01GM127594-02
- **Recipient organization:** UNIVERSITY OF ARIZONA
- **Principal Investigator:** STEVEN D SCHWARTZ
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $297,920
- **Award type:** 5
- **Project period:** 2019-01-01 → 2022-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9839628

## Citation

> US National Institutes of Health, RePORTER application 9839628, Rapid protein dynamics and catalysis: modulation by laboratory evolution, designed mutation, and protein control of electric field environment (5R01GM127594-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9839628. Licensed CC0.

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