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GTPases and their Regulation

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Guanine nucleotide-binding proteins (G-proteins), also known as GTPases, are a superfamily of proteins that regulate many cellular processes, such as cell signaling, vesicular transport, and the regulation of cell shape and motility. Mutation or dysfunction of these proteins can lead to disease. There are around 40,000 known G-proteins that can broadly be classified into two groups ‒ small G-proteins consisting of a single domain and large multi-domain G-proteins.
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GTPases and their Regulation

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Related Experiment Video

Updated: May 11, 2026

Comparing the Affinity of GTPase-binding Proteins using Competition Assays

Comparing the Affinity of GTPase-binding Proteins using Competition Assays

Published on: October 8, 2015

G domain dimerization controls dynamin's assembly-stimulated GTPase activity.

Joshua S Chappie¹, Sharmistha Acharya, Marilyn Leonard

¹Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, NIH, Bethesda, Maryland 20892, USA.

|April 30, 2010

Summary

Dynamin, a GTPase crucial for cell membrane fission, was studied using a crystal structure. This reveals how dynamin

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Area of Science:

Biochemistry
Molecular Biology
Cell Biology

Background:

Dynamin is an atypical GTPase essential for membrane fission during clathrin-mediated endocytosis.
The precise mechanisms governing dynamin's basal and assembly-stimulated GTP hydrolysis remain largely unknown.
The GTPase effector domain (GED) is known to indirectly influence dynamin's GTPase activity.

Purpose of the Study:

To elucidate the structural basis of dynamin's GTPase activity and its regulation.
To understand the mechanism of assembly-stimulated GTP hydrolysis in dynamin.
To provide insights into dynamin-catalyzed membrane fission.

Main Methods:

Determined the 2.0 Å resolution crystal structure of a human dynamin 1-derived minimal GTPase-GED fusion protein.
Utilized the transition state mimic GDP.AlF(4)(-) to stabilize the dimeric form of the protein.
Performed structural comparisons with the rat dynamin G domain.

Main Results:

The crystal structure revealed the dimeric state of the GTPase-GED fusion protein in the presence of GDP.AlF(4)(-).
The structure elucidated dynamin's catalytic machinery and demonstrated G domain dimerization as the mechanism for assembly-stimulated GTP hydrolysis.
A sodium ion was identified in the active site, suggesting its role in stabilizing the transition state in the absence of an arginine finger.

Conclusions:

The study provides a structural explanation for how dynamin achieves assembly-stimulated GTP hydrolysis via G domain dimerization.
The findings highlight the role of a cation in the active site for transition state stabilization.
The presented structure offers significant insights into the molecular mechanisms underlying dynamin-mediated membrane fission.