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Related Experiment Video

Updated: Jul 2, 2025

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050

Published on: May 13, 2020

Perturbative diffraction methods resolve a conformational switch that facilitates a two-step enzymatic mechanism.

Jack B Greisman¹, Kevin M Dalton¹, Dennis E Brookner¹

¹Department of Molecular & Cellular Biology, Harvard University, Cambridge, MA 02138.

Proceedings of the National Academy of Sciences of the United States of America

|February 22, 2024

Summary

Protein dynamics are crucial for enzyme catalysis. This study reveals how Escherichia coli dihydrofolate reductase (DHFR) uses conformational dynamics to facilitate proton and hydride transfer, enhancing catalytic efficiency.

Keywords:

DHFR X-ray crystallography allostery enzyme catalysis protein dynamics

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Last Updated: Jul 2, 2025

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050

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Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes

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Published on: January 16, 2016

Area of Science:

Biochemistry
Structural Biology
Enzymology

Background:

Enzymes catalyze reactions by precisely positioning substrates and modulating transition-state energy.
The role of protein conformational dynamics in enzyme catalysis is poorly understood due to experimental limitations.
Escherichia coli dihydrofolate reductase (DHFR) is a model enzyme for studying protein dynamics in catalysis.

Purpose of the Study:

To investigate the role of conformational dynamics in the catalytic mechanism of Escherichia coli dihydrofolate reductase (DHFR).
To understand how DHFR regulates its active site environment to facilitate proton and hydride transfer during catalysis.

Main Methods:

Utilized X-ray diffraction experiments with ligand, temperature, and electric-field perturbations.
Mapped the conformational dynamics of the DHFR Michaelis complex.
Resolved coupled global and local motions within the enzyme.

Main Results:

Identified coupled global and local motions in the DHFR Michaelis complex.
Demonstrated that these motions are engaged by the protonated substrate.
Showed that substrate engagement promotes efficient catalysis.

Conclusions:

Protein conformational dynamics are integral to enzyme catalysis, not just passive facilitators.
Pre-existing enzyme dynamics enable intermediates to drive electrostatic reorganization for subsequent catalytic steps.
This suggests a fundamental design principle for multistep enzymes involving dynamics-driven catalysis.