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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...

Induced-fit Model

Induced-fit Model

Most chemical reactions in cells require enzymes—biological catalysts that speed up the reaction without being consumed or permanently changed. They reduce the activation energy needed to convert the reactants into products. Enzymes are proteins, that usually work by binding to a substrate—a reactant molecule that they act upon.
Enzymes exhibit substrate specificity, meaning that they can only bind to certain substrates. This is mainly determined by the shape and chemical characteristics of...

Enzyme Kinetics

Enzyme Kinetics

Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...

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: May 20, 2026

Luciferase Complementation Imaging Assay in Nicotiana benthamiana Leaves for Transiently Determining Protein-protein Interaction Dynamics

Luciferase Complementation Imaging Assay in Nicotiana benthamiana Leaves for Transiently Determining Protein-protein Interaction Dynamics

Published on: November 20, 2017

Substrate cooperativity in marine luciferases.

George Tzertzinis¹, Ezra Schildkraut, Ira Schildkraut

¹New England Biolabs, Inc., Ipswich, Massachusetts, United States of America.

|July 7, 2012

Summary

Marine luciferases from Gaussia (GLuc) and Metridia (MLuc) exhibit cooperative light production with substrate concentration. This contrasts with Renilla (RLuc) and Cypridina (CLuc) luciferases, showing linear responses.

Area of Science:

Biochemistry
Molecular Biology
Enzymology

Background:

Marine luciferases are valuable tools for studying gene regulation and developing bioluminescent assays.
Limited information exists regarding the catalytic properties of marine luciferases concerning substrate concentration.
Understanding enzyme kinetics is crucial for optimizing bioluminescent assay performance.

Purpose of the Study:

To investigate and compare the substrate concentration-dependent catalytic properties of marine luciferases.
To elucidate the kinetic mechanisms of Gaussia princeps (GLuc) and Metridia longa (MLuc) luciferases.
To contrast the kinetic behavior of GLuc and MLuc with Renilla reniformis (RLuc) and Cypridina noctiluca (CLuc) luciferases.

Main Methods:

Enzyme kinetics assays were performed to measure light production across varying substrate concentrations for GLuc, MLuc, RLuc, and CLuc.

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Monitoring Endoplasmic Reticulum Calcium Homeostasis Using a Gaussia Luciferase SERCaMP

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Analysis of light output rates in response to a 10-fold decrease in luciferin substrate concentration.

Characterization of cooperative and linear kinetic models for marine luciferases.

Main Results:

Gaussia princeps (GLuc) and Metridia longa (MLuc) luciferases displayed a surprising cooperative light production mechanism.
A 10-fold decrease in substrate concentration resulted in a 1000-fold decrease in light output for GLuc and MLuc.
Renilla reniformis (RLuc) and Cypridina noctiluca (CLuc) luciferases exhibited a linear relationship between substrate concentration and light production.

Conclusions:

The cooperative kinetics of GLuc and MLuc are likely attributed to allosteric interactions between their catalytic domains.
The distinct kinetic profiles of marine luciferases have significant implications for bioluminescent assay design and interpretation.
Further research into the allosteric mechanisms of marine luciferases could lead to improved reporter systems.