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

Enzymes02:34

Enzymes

97.6K
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...
97.6K
Introduction to Mechanisms of Enzyme Catalysis01:13

Introduction to Mechanisms of Enzyme Catalysis

11.7K
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...
11.7K
Induced-fit Model01:13

Induced-fit Model

92.2K
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...
92.2K

You might also read

Related Articles

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

Sort by
Same author

Mavacamten shows broad benefit in human and mouse models of MYBPC3-related hypertrophic cardiomyopathy.

Nature cardiovascular research·2026
Same author

Fibronectin-induced overactivation of α<sub>V</sub>β<sub>3</sub>-PI3K-PIP3-PDK1-ILK signaling drives aortic disease in Marfan syndrome.

Nature communications·2026
Same author

Reclone: a global research community building equitable access to reagents.

Nature methods·2026
Same author

Functional conservation with mechanistic drift: AMP activation in the evolution of archaeal sugar kinases.

Molecular biology and evolution·2026
Same author

Titin cleavage in living cardiomyocytes induces sarcomere disassembly but does not trigger cell proliferation.

The Journal of biological chemistry·2026
Same author

Mending the Achilles heels of titin in cardiac and musculoskeletal disease.

Biophysical reviews·2026

Related Experiment Video

Updated: Apr 15, 2026

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
08:34

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy

Published on: February 5, 2020

7.3K

Identifying sequential substrate binding at the single-molecule level by enzyme mechanical stabilization.

Jaime Andrés Rivas-Pardo1,2, Jorge Alegre-Cebollada1, César A Ramírez-Sarmiento2

  • 1†Department of Biological Sciences, Columbia University, Northwest Corner Building, 550 West 120 Street, New York, New York 10027, United States.

ACS Nano
|April 5, 2015
PubMed
Summary

Single-molecule force spectroscopy reveals how substrate binding alters the unfolding energy of Thermococcus litoralis glucokinase. Enzyme structural changes and domain closure were observed upon sequential substrate addition.

Keywords:
force−extensionmechanical clampmechanical intermediatesingle-molecule force spectroscopysubstrate stabilization

More Related Videos

Investigating Receptor-ligand Systems of the Cellulosome with AFM-based Single-molecule Force Spectroscopy
11:34

Investigating Receptor-ligand Systems of the Cellulosome with AFM-based Single-molecule Force Spectroscopy

Published on: December 20, 2013

7.9K
Dissecting Mechanoenzymatic Properties of Processive Myosins with Ultrafast Force-Clamp Spectroscopy
09:38

Dissecting Mechanoenzymatic Properties of Processive Myosins with Ultrafast Force-Clamp Spectroscopy

Published on: July 1, 2021

1.7K

Related Experiment Videos

Last Updated: Apr 15, 2026

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
08:34

OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy

Published on: February 5, 2020

7.3K
Investigating Receptor-ligand Systems of the Cellulosome with AFM-based Single-molecule Force Spectroscopy
11:34

Investigating Receptor-ligand Systems of the Cellulosome with AFM-based Single-molecule Force Spectroscopy

Published on: December 20, 2013

7.9K
Dissecting Mechanoenzymatic Properties of Processive Myosins with Ultrafast Force-Clamp Spectroscopy
09:38

Dissecting Mechanoenzymatic Properties of Processive Myosins with Ultrafast Force-Clamp Spectroscopy

Published on: July 1, 2021

1.7K

Area of Science:

  • Biochemistry
  • Structural Biology
  • Biophysics

Background:

  • Enzyme-substrate binding is intrinsically dynamic, influencing protein structure and energy landscapes.
  • Understanding these conformational changes is key to elucidating enzyme mechanisms.

Purpose of the Study:

  • To investigate the conformational transitions of Thermococcus litoralis glucokinase (TlGK) upon sequential substrate binding.
  • To characterize the mechanical unfolding of TlGK using single-molecule force spectroscopy (SMFS).

Main Methods:

  • Single-molecule force spectroscopy (SMFS) was employed to probe TlGK unfolding.
  • The enzyme's mechanical unfolding was analyzed in the absence and presence of substrates (Mg·ADP and ATP).

Main Results:

  • Substrate binding, particularly Mg·ADP, stabilized an unfolding intermediate (intermediate 1).
  • A novel unfolding intermediate (intermediate 1*) emerged upon Mg·ADP binding.
  • Both intermediates showed increased unfolding forces with the addition of the second substrate, indicating domain closure.

Conclusions:

  • SMFS effectively characterizes enzyme conformational changes induced by sequential substrate binding.
  • The study provides insights into the dynamic binding mechanism of TlGK.
  • This approach expands the toolkit for studying multi-ligand enzyme interactions.