# Structural and Dynamic Mechanisms in Classical Protein Allostery

> **NIH NIH R01** · UNIV OF NORTH CAROLINA CHAPEL HILL · 2021 · $348,273

## Abstract

Abstract
Allosteric regulation of protein activity is a physico-mechanical phenomenon that underlies the coordination of
cellular events throughout biology. Signal transduction, metabolism, and other essential cellular processes are
completely reliant on the executions of conformational and dynamic changes that enable allosteric proteins to
communicate between distant sites. To understand such biological mechanisms – and by extension to
understand how to rationally alter cellular processes, either with drugs or protein engineering – is to understand
this fundamental problem of how allosteric regulation works. Yet, even though allosteric regulation has been
recognized for decades and despite the recent realization that dynamics contributes to allostery, our
understanding of allosteric mechanism is still at a rudimentary level. One limitation has been that the roles of
dynamics in allostery have been drawn from just a few systems, most of which lack the classic indicators of
functional allostery. Another limitation is that gaining accurate information on functional dynamics is
experimentally challenging. To identify basic working principles of allostery, mechanisms of allosteric behavior
must be observed in proteins that are “strongly allosteric”, where allosteric movements and signatures will be
more easily identified. In the long term, knowledge of allosteric mechanism will enhance protein research in
general and have a huge positive impact on design of allosteric drugs and allosteric proteins. The focus of this
work will be on the allosteric enzyme chorismate mutase (CM). By all considerations, this enzyme appears to be
ideal for high-resolution dissective studies of its allosteric mechanisms. CM is a canonical allosteric enzyme as
evidenced by a number of characteristics: it is a symmetric dimer with active sites separated by 40 Å; it
undergoes T-to-R conformational transitions; it exhibits homotropic allostery (Hill coefficient = 1.6); and it exhibits
heterotropic allostery with small molecule effectors that modulate activity up (by Trp) or down (by Tyr). CM is 60
kDa which makes it amenable to solution NMR studies, and it is extremely soluble and durable and yields
outstanding quality NMR spectra. The rich allosteric characteristics of CM will allow classical allostery to be
examined experimentally using NMR and other biochemical and biophysical methods (including computations)
in unprecedented detail. In this proposal, Aims 1 and 2 employ NMR, computational methods, and chemical
synthesis to characterize the structural and dynamic features of apo and liganded states of CM in solution. The
responses of CM to binding effectors and a transition state analog will be monitored, all towards the goal of
identification of mechanisms of heterotropic long-range communication. Aim 3 is focused on extending a novel
labeling methodology for monitoring mechanisms of homotropic allostery. “Click” chemistry will be used to
covalently and specifically tether...

## Key facts

- **NIH application ID:** 10216306
- **Project number:** 5R01GM127698-03
- **Recipient organization:** UNIV OF NORTH CAROLINA CHAPEL HILL
- **Principal Investigator:** Andrew L Lee
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $348,273
- **Award type:** 5
- **Project period:** 2019-09-20 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10216306

## Citation

> US National Institutes of Health, RePORTER application 10216306, Structural and Dynamic Mechanisms in Classical Protein Allostery (5R01GM127698-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10216306. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*