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

Introduction to Mechanisms of Enzyme Catalysis01:13

Introduction to Mechanisms of Enzyme Catalysis

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 a mild...
Introduction to Mechanisms of Enzyme Catalysis01:13

Introduction to Mechanisms of Enzyme Catalysis

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 a mild...
Allosteric Proteins-ATCase01:19

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,...
Catalytically Perfect Enzymes01:07

Catalytically Perfect Enzymes

The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.
Ligand Binding and Linkage00:49

Ligand Binding and Linkage

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 the...
Mechanical Protein Functions01:58

Mechanical Protein Functions

Proteins perform many mechanical functions in a cell. These proteins can be classified into two general categories- proteins that generate mechanical forces and proteins that are subjected to mechanical forces. Proteins providing mechanical support to the structure of the cell, such as keratin, are subjected to mechanical force, whereas proteins involved in cell movement and transport of molecules across cell membranes, such as an ion pump, are examples of generating mechanical force. 

You might also read

Related Articles

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

Sort by
Same author

When Brownian Motion Meets Clinical Laboratory Automation: A DLS-Inspired Autocorrelation Function for Characterizing Workflow Performance in Sample Processing.

Diagnostics (Basel, Switzerland)·2026
Same author

Origin of the High Variability in Sol-Gel Phase Transitions: The Agar Gelation Model.

Gels (Basel, Switzerland)·2026
Same author

Permanent weakness and myopathy in hypokalemic periodic paralysis.

Acta myologica : myopathies and cardiomyopathies : official journal of the Mediterranean Society of Myology·2026
Same author

Dialysis Unit Differences in Post-Dialysis Fatigue Prevalence and Characteristics and Time to Recovery After Dialysis.

Hemodialysis international. International Symposium on Home Hemodialysis·2026
Same author

Association Between Postdialysis Fatigue and Number of Comorbidities and Number and Prevalence of Symptoms.

Hemodialysis international. International Symposium on Home Hemodialysis·2026
Same author

Impact of Stenting with Angioplasty and MTICI 2c-3 Recanalization On Outcome in Acute MCA Occlusion with Underlying Stenosis.

Clinical neuroradiology·2025

Related Experiment Video

Updated: Jul 5, 2026

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

Mechanistic studies on ADAMTS13 catalysis.

Enrico Di Stasio1, Stefano Lancellotti, Flora Peyvandi

  • 1Institute of Biochemistry and Clinical Biochemistry, Catholic University School of Medicine, Rome, Italy.

Biophysical Journal
|May 27, 2008
PubMed
Summary

The metalloprotease ADAMTS13 cleaves von Willebrand factor (VWF) to prevent dangerous blood clots. This study reveals how temperature and pH affect ADAMTS13 activity and identifies key VWF regions crucial for substrate recognition and cleavage.

More Related Videos

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

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050
11:27

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050

Published on: May 13, 2020

Related Experiment Videos

Last Updated: Jul 5, 2026

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

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

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050
11:27

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050

Published on: May 13, 2020

Area of Science:

  • Biochemistry
  • Enzymology
  • Proteolysis

Background:

  • ADAMTS13 (a disintegrin-like and metalloprotease with thrombospondin type I repeats) is essential for cleaving von Willebrand factor (VWF).
  • Dysfunctional ADAMTS13 activity leads to the accumulation of ultra-large VWF multimers, associated with thrombotic thrombocytopenic purpura.

Purpose of the Study:

  • To investigate the catalytic mechanism and substrate recognition of ADAMTS13.
  • To determine the influence of temperature and pH on ADAMTS13 activity.
  • To elucidate the structural basis for VWF recognition and cleavage by ADAMTS13.

Main Methods:

  • Utilized a fluorescence quenching method (FRETS method) to monitor the hydrolysis of a VWF analog (FRETS-VWF73).
  • Assessed enzyme kinetics across a range of temperatures (15–45 °C) and pH values (4.5–10.5).
  • Employed molecular modeling to visualize substrate-enzyme interactions.

Main Results:

  • Catalysis was influenced by two ionizable groups with specific pK(a) values and ionization enthalpies.
  • Negative activation entropy suggests an ordered transition state during substrate hydrolysis.
  • Identified a 'hot spot' in the VWF A2 sequence (Asp1653-Thr1656-Glu1660-Asp1663) critical for ADAMTS13 binding and allosteric regulation.
  • The VWF cleavage product (Met1606-Arg1668) acts as a mixed-type inhibitor.
  • Enzyme activity is progressively inhibited at pH values above 8.50 due to interactions with basic residues.

Conclusions:

  • ADAMTS13 activity is finely tuned by environmental factors like pH and temperature.
  • Specific acidic residues and the overall conformation of ADAMTS13 are crucial for efficient VWF cleavage.
  • Understanding these interactions provides insights into ADAMTS13 function and potential therapeutic strategies.