Related Experiment Video
Updated: Dec 29, 2025
08:03
Study of Protein Dynamics via Neutron Spin Echo Spectroscopy
Published on: April 13, 2022
2.5K
Nanosecond-Timescale Dynamics and Conformational Heterogeneity in Human GCK Regulation and Disease
Shawn M Sternisha1, A Carl Whittington2, Juliana A Martinez Fiesco1
1Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida.
Biophysical Journal
|February 6, 2020
Summary
Human glucokinase (GCK) dynamics were explored using advanced spectroscopy. Faster motions reveal how glucose binding and mutations alter enzyme conformations, explaining cooperativity.
Area of Science:
- Biochemistry
- Structural Biology
- Enzymology
Background:
- Human glucokinase (GCK) exhibits allosteric behavior driven by intrinsic protein dynamics.
- Understanding GCK's dynamics is crucial for modeling enzyme cooperativity.
- Faster protein motions (nanosecond timescale) in GCK remain largely unexplored.
Purpose of the Study:
- To map nanosecond-timescale dynamics and structural heterogeneity in human glucokinase.
- To investigate how glucose binding and disease-associated mutations affect GCK dynamics.
- To develop a comprehensive dynamical framework for GCK cooperativity.
Main Methods:
- Incorporation of unnatural amino acids.
- Time-resolved fluorescence spectroscopy.
- Fluorine-19 nuclear magnetic resonance (19F NMR) spectroscopy.
Main Results:
- A probe in an intrinsically disordered loop of GCK samples multiple conformations in the unliganded state.
- Glucose binding and cooperative-disrupting mutations alter the population and number of these conformational states.
- Nanosecond dynamics complement known millisecond dynamics to explain GCK cooperativity.
Conclusions:
- Nanosecond dynamics are integral to the allosteric mechanism of human glucokinase.
- Alterations in conformational states correlate with changes in GCK cooperativity.
- This study provides a dynamical framework for understanding GCK function and disease-associated variants.
More Related Videos
Related Concept Videos
02:14GTPases and their Regulation
9.6K
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,...
9.6K
00:52¹H NMR of Conformationally Flexible Molecules: Temporal Resolution
1.2K
At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the...
1.2K
01:22Activation and Inactivation of G Proteins
10.5K
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...
10.5K