Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Induced-fit Model01:13

Induced-fit Model

92.0K
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...
92.0K
Enzymes02:34

Enzymes

97.2K
Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
97.2K
Allosteric Proteins-ATCase01:19

Allosteric Proteins-ATCase

6.9K
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...
6.9K
Introduction to Mechanisms of Enzyme Catalysis01:13

Introduction to Mechanisms of Enzyme Catalysis

11.5K
For many years, scientists thought that enzyme-substrate binding took place in a simple "lock-and-key" fashion. This model stated that the enzyme and substrate fit together perfectly in one instantaneous step. However, current research supports a more refined view scientists call induced fit. The induced-fit model expands upon the lock-and-key model by describing a more dynamic interaction between enzyme and substrate. As the enzyme and substrate come together, their interaction causes...
11.5K
Introduction to Mechanisms of Enzyme Catalysis01:13

Introduction to Mechanisms of Enzyme Catalysis

10.2K
10.2K
Ligand Binding and Linkage00:49

Ligand Binding and Linkage

6.0K
Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked.  In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...
6.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Deciphering AMP deaminase-2 structure, activators and regulators underpinning cellular function in human fructose and nucleotide metabolism.

bioRxiv : the preprint server for biology·2026
Same author

Glycoconjugate diversification in <i>Campylobacter concisus</i> is determined by two glycosyltransferases.

bioRxiv : the preprint server for biology·2026
Same author

Fructose: metabolic signal and modern hazard.

Nature metabolism·2026
Same author

Detergent Exchange from Lipid Nanoparticles into Detergent Micelles Unlocks a Tool for Biochemical and Kinetic Characterization of Membrane Proteins.

Biochemistry·2026
Same author

Detergent exchange from lipid nanoparticles into detergent micelles unlocks a tool for biochemical and kinetic characterization of membrane proteins.

bioRxiv : the preprint server for biology·2026
Same author

Identification of a common ketohexokinase-dependent link driving alcohol intake and alcohol-associated liver disease in mice.

Nature metabolism·2025
Same journal

Switching Site Selectivity in Alkoxyamine Hydration: From Lone-Pair Direction to Solvent Network Dominance.

Journal of the American Chemical Society·2026
Same journal

A Topotactic Leap: 2D Layers to 3D Large-Pore Zeolite.

Journal of the American Chemical Society·2026
Same journal

Enhanced Hydrogen Evolution over Single-Atom Catalysts via Electrostatic Polarization in Contact-electro-catalysis.

Journal of the American Chemical Society·2026
Same journal

Tumor Acidity-Activatable Ionizable Lipid Nanoparticles for Selective Oncolytic Therapy.

Journal of the American Chemical Society·2026
Same journal

Alternating Magnetic Field Promotes Ammonia Cracking by Disrupting the Sabatier Limitation of Ruthenium Catalytic Species.

Journal of the American Chemical Society·2026
Same journal

Bulk Ferromagnetic Icosahedral Quasicrystals without Rapid Quenching.

Journal of the American Chemical Society·2026
See all related articles

Related Experiment Video

Updated: Apr 1, 2026

Defining Substrate Specificities for Lipase and Phospholipase Candidates
08:59

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

15.7K

Enzyme Substrate Specificity Conferred by Distinct Conformational Pathways.

Florencia Rago1, Daniel Saltzberg2, Karen N Allen1,3

  • 1Program in Biochemistry and Molecular Biology (BMB), Boston University , Boston, Massachusetts 02215, United States.

Journal of the American Chemical Society
|October 7, 2015
PubMed
Summary
This summary is machine-generated.

Enzyme conformational changes dictate substrate specificity. Aldolase uses different conformations to distinguish between fructose 1-phosphate and fructose 1,6-bisphosphate, linking conformational shifts to catalysis.

More Related Videos

Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes
09:42

Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes

Published on: January 16, 2016

9.5K
Modeling an Enzyme Active Site using Molecular Visualization Freeware
14:37

Modeling an Enzyme Active Site using Molecular Visualization Freeware

Published on: December 25, 2021

11.9K

Related Experiment Videos

Last Updated: Apr 1, 2026

Defining Substrate Specificities for Lipase and Phospholipase Candidates
08:59

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

15.7K
Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes
09:42

Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes

Published on: January 16, 2016

9.5K
Modeling an Enzyme Active Site using Molecular Visualization Freeware
14:37

Modeling an Enzyme Active Site using Molecular Visualization Freeware

Published on: December 25, 2021

11.9K

Area of Science:

  • Biochemistry
  • Enzyme kinetics
  • Protein conformational dynamics

Background:

  • Enzyme catalysis relies on substrate recognition, but direct evidence linking enzyme conformational changes to substrate selectivity is limited.
  • Aldolase, a key glycolytic enzyme, must differentiate between fructose 1-phosphate and fructose 1,6-bisphosphate, making it an ideal model for studying substrate discrimination.
  • Isozyme-specific residues (ISRs) distant from the active site, particularly in surface alpha-helices and the carboxyl-terminal region (CTR), are known to mediate kinetic distinctions.

Purpose of the Study:

  • To investigate the hypothesis that enzyme conformational changes underlie substrate selectivity in aldolase.
  • To explore the role of specific regions, including surface alpha-helices and the CTR, in mediating these conformational shifts.
  • To establish a link between observed conformational dynamics and the catalytic process.

Main Methods:

  • Creation of single surface-cysteine variants of aldolase for site-specific labeling.
  • Labeling variants with environmentally sensitive fluorophores.
  • Monitoring conformational changes using fluorescence emission spectrophotometry and stopped-flow fluorescence spectrophotometry.

Main Results:

  • Fluorescence spectra of labeled aldolase variants differed in the presence of saturating amounts of fructose 1-phosphate versus fructose 1,6-bisphosphate, indicating distinct conformations.
  • Substrate concentration-dependent changes in fluorescence spectra confirmed binding events.
  • Stopped-flow fluorescence measurements revealed that the rate of conformational changes occurred on the same timescale as catalysis.

Conclusions:

  • Aldolase exhibits substrate-specific conformational changes.
  • These conformational dynamics are linked to the enzyme's catalytic cycle and substrate recognition.
  • Conformational changes represent a potential common mechanism for substrate specificity in enzymes with multiple substrates.