Related Experiment Video
Updated: Apr 25, 2026

13:51
Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
Published on: November 11, 2018
9.4K
What vibrations tell us about GTPases.
Biological Chemistry
|August 26, 2014
Summary
Time-resolved Fourier transform infrared (FTIR) spectroscopy reveals the atomic details of GTP hydrolysis catalyzed by small GTPases and their activating proteins. This method illuminates disease-related mechanisms and catalytic processes at the membrane.
Area of Science:
- Biochemistry and Molecular Biology
- Spectroscopy
- Cell Signaling
Background:
- Small GTPases and their GTPase-activating proteins (GAPs) regulate crucial cellular processes like signal transduction and transport.
- Dysfunctional GTP hydrolysis by these proteins is implicated in severe diseases, including cancer.
- Understanding the precise catalytic mechanisms is vital for therapeutic development.
Purpose of the Study:
- To review the application of time-resolved Fourier transform infrared (FTIR) spectroscopy for elucidating GTP hydrolysis mechanisms.
- To detail how FTIR provides atomic-level insights into GTPase-catalyzed reactions.
- To explore GTPase function under near-physiological membrane conditions.
Main Methods:
- Time-resolved Fourier transform infrared (FTIR) spectroscopy to resolve GTPase reactions with high spatiotemporal resolution.
- Attenuated total reflection (ATR) technique to study GTPases at membrane interfaces.
- Comparative analysis of different small GTPases to identify conserved and variable catalytic features.
Main Results:
- FTIR successfully resolved the intrinsic and GAP-catalyzed GTPase reaction of Ras, revealing the order of catalytic events.
- Detailed atomic insights into active site completion during catalysis were obtained.
- Conservation and variation in catalytic mechanisms across different small GTPases were identified.
Conclusions:
- Time-resolved FTIR spectroscopy is a powerful tool for dissecting GTPase catalytic mechanisms at atomic resolution.
- The study provides a deeper understanding of GTP hydrolysis, relevant to cellular signaling and disease.
- ATR-FTIR extends mechanistic studies to more physiologically relevant membrane environments.
More Related Videos
Related Concept Videos
GTPases and their Regulation
7.9K
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,...
Large G-proteins,...
7.9K
GTPases and their Regulation
2.4K
2.4K
Activation and Inactivation of G Proteins
8.8K
Heterotrimeric G proteins are guanine nucleotide-binding proteins. As the name suggests, heterotrimeric G proteins are composed of three subunits: alpha, beta, and gamma. They remain GDP-bound or GTP-bound inside the cells and switch between inactive/active states. The Gα subunit possesses the nucleotide-binding pocket that binds guanine nucleotides and switches between GDP or GTP-bound states. In contrast, the Gꞵ and Gγ subunits are always bound together with high...
8.8K
Small GTPases - Ras and Rho
4.4K
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:
Three regulatory proteins control their activity:
4.4K
Coat Assembly and GTPases
3.5K
Vesicles incorporate different coat protein subunits in different cell locations, which changes the properties of the coat, such as the shape and geometry of the transport vesicles. Thus, vesicle coat proteins also play a significant role in cargo selection.
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
3.5K
Mechanical Protein Functions
4.4K
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.
4.4K

