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Allosteric Regulation

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...

Allosteric Regulation

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...

Cooperative Allosteric Transitions

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...

Cooperative Allosteric Transitions

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...

Cooperative Allosteric Transitions

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...

Allosteric Proteins-ATCase

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...

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Related Experiment Video

Updated: Jul 3, 2026

Assaying the Kinase Activity of LRRK2 in vitro

Assaying the Kinase Activity of LRRK2 in vitro

Published on: January 18, 2012

Allosteric coupling in pyruvate dehydrogenase kinase 2.

Alla Klyuyeva¹, Alina Tuganova, Kirill M Popov

¹Department of Biochemistry and Molecular Genetics, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.

|July 17, 2008

Summary

Mitochondrial pyruvate dehydrogenase kinase 2 (PDHK2) mutations near its active site disrupt regulation by dichloroacetate (DCA) and other metabolites. This uncouples PDHK2

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Area of Science:

Biochemistry
Molecular Biology
Enzyme kinetics

Background:

Mitochondrial pyruvate dehydrogenase kinase 2 (PDHK2) regulates pyruvate metabolism by phosphorylating the pyruvate dehydrogenase complex (PDC).
PDHK2 activity is modulated by various metabolites, including pyruvate, NAD+, NADH, CoA, and acetyl-CoA, binding to distinct allosteric sites.
Pyruvate and its analogue dichloroacetate (DCA) inhibit PDHK2 via a specific binding site, distinct from the metabolite-binding site.

Purpose of the Study:

To investigate the structural basis of allosteric regulation in PDHK2.
To determine the role of specific amino acid residues near the active site in mediating PDHK2 regulation.
To explore how mutations affect the communication between PDHK2's active and allosteric sites.

Main Methods:

Site-directed mutagenesis of PDHK2 at residues R250, T302, and Y320.
Enzyme activity assays to assess inhibition by DCA and other metabolites.
Analysis of structural data to infer conformational changes and regulatory mechanisms.

Main Results:

Mutations R250, T302, and Y320 confer resistance to DCA inhibition, uncoupling the active site from the allosteric DCA-binding site.
Substitutions at R250 and T302 also partially or fully uncouple the L2-binding site for other metabolites.
These residues stabilize the conformational states of an ATP-binding lid, suggesting its role in allosteric regulation.

Conclusions:

The mobility of the ATP lid, stabilized by R250, T302, and Y320, is crucial for PDHK2 allosteric regulation.
These findings reveal a conformational switch mechanism for communication between PDHK2's active and allosteric sites.
Understanding these regulatory mechanisms provides insights into metabolic control and potential therapeutic targets.