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Related Concept Videos

GTPases and their Regulation02:14

GTPases and their Regulation

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.
Large G-proteins, also known...
GTPases and their Regulation02:14

GTPases and their Regulation

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.
Large G-proteins, also known...
Cryo-electron Microscopy01:28

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
Small GTPases - Ras and Rho01:24

Small GTPases - Ras and Rho

Ras and Rho are small monomeric GTPases that act downstream of receptor tyrosine kinase (RTK) and regulate various cellular processes. These GTPases switch between active and inactive states by binding to guanine nucleotides.
Three regulatory proteins control their activity:
Mechanical Protein Functions01:58

Mechanical Protein Functions

Proteins perform many mechanical functions in a cell. These proteins can be classified into two general categories- proteins that generate mechanical forces and proteins that are subjected to mechanical forces. Proteins providing mechanical support to the structure of the cell, such as keratin, are subjected to mechanical force, whereas proteins involved in cell movement and transport of molecules across cell membranes, such as an ion pump, are examples of generating mechanical force. 

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Related Experiment Video

Updated: May 16, 2026

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy
08:27

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy

Published on: January 7, 2019

Protein Engineering-Enabled Cryo-EM Investigation of Small GTPases.

Zhengshan Hu1, Unnatiben Rajeshbhai Patel1, Eliezra Glasser1

  • 1Perlmutter Cancer Center, New York University Langone Health, New York, NY, USA.

Journal of Molecular Biology
|May 14, 2026
PubMed
Summary

Researchers developed a novel "RAS-lollipop" cryo-electron microscopy (cryo-EM) method to visualize small GTPases like NRAS. This technique overcomes crystal structure limitations, revealing unperturbed conformations and binding sites for drug discovery.

Keywords:
AlphaFold predictionhelix plasticitylinker optimizationscaffold-assisted structure determination

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Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
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Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay

Published on: November 11, 2018

A Pipeline to Investigate the Structures and Signaling Pathways of Sphingosine 1-Phosphate Receptors
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A Pipeline to Investigate the Structures and Signaling Pathways of Sphingosine 1-Phosphate Receptors

Published on: June 8, 2022

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Last Updated: May 16, 2026

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy
08:27

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy

Published on: January 7, 2019

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
13:51

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay

Published on: November 11, 2018

A Pipeline to Investigate the Structures and Signaling Pathways of Sphingosine 1-Phosphate Receptors
12:27

A Pipeline to Investigate the Structures and Signaling Pathways of Sphingosine 1-Phosphate Receptors

Published on: June 8, 2022

Area of Science:

  • Structural Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Small GTPases are crucial for cellular signaling but challenging to study structurally due to their small size.
  • X-ray crystallography of small GTPases is hindered by crystal lattice contacts affecting key switch regions.

Purpose of the Study:

  • To develop a protein-engineering strategy for cryo-EM analysis of small soluble proteins, specifically RAS.
  • To determine the unperturbed structure of NRAS using cryo-EM and identify potential drug binding sites.

Main Methods:

  • Engineered a "RAS-lollipop" complex by fusing NRAS to BRIL and an anti-BRIL Fab fragment.
  • Utilized single-particle cryo-electron microscopy (cryo-EM) for structural determination.
  • Characterized the binding site and properties of an NRAS-specific monobody.

Main Results:

  • Determined the cryo-EM structure of NRAS, revealing novel conformations of switch regions and the α5 helix.
  • Identified the binding site of an NRAS-specific monobody.
  • Discovered a surfactant-like property of the monobody that improves cryo-EM grid particle orientation.

Conclusions:

  • The "RAS-lollipop" approach provides a platform for visualizing small GTPases and other small proteins with minimal surface perturbation.
  • This method enables structural studies of proteins previously challenging for crystallography, like NRAS.
  • The findings facilitate understanding of RAS signaling and the development of targeted therapeutics.