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

Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass. One common type of ionization, known as electron ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave behind a...
High-Resolution Mass Spectrometry (HRMS)01:15

High-Resolution Mass Spectrometry (HRMS)

The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For example, the mass of helium...
Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
Mass Spectrometry: Isotope Effect01:13

Mass Spectrometry: Isotope Effect

Most elements exist in nature as a mixture of isotopes. The isotopes differ in weight due to their respective number of neutrons. The molecular weight of a molecule is different depending on the specific isotope of its elements involved. As a result, the mass spectrum of the molecule exhibits peaks from the same fragment at multiple positions. The positions of these mass signals depend on the mass differences between isotopes. Furthermore, the intensity of these signals is dependent on the...
Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...

You might also read

Related Articles

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

Sort by
Same author

On the relationship between efficiency and cooperativity in multi-subunit molecular machines.

Protein science : a publication of the Protein Society·2026
Same author

Editorial: Ubiquitin proteasome system (UPS) and ubiquitin-independent proteasome-mediated proteolysis (UIPP) crosstalk in development and disease.

Frontiers in cell and developmental biology·2025
Same author

Cleaving Expectations: A Review of Proteasome Functional and Catalytic Diversity.

Biomolecules·2025
Same author

Merging different allosteric mechanisms: The case of <i>Escherichia coli</i> glutathione reductase.

Proceedings of the National Academy of Sciences of the United States of America·2025
Same author

Inhibition of tau aggregation by the CCT3 and CCT7 apical domains.

Protein science : a publication of the Protein Society·2025
Same author

Tracking proteasome degradation: A cross-organ analysis via intact degradomics mass spectrometry.

Proceedings of the National Academy of Sciences of the United States of America·2025

Related Experiment Video

Updated: May 12, 2026

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
10:01

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

Published on: November 28, 2017

Allosteric mechanisms can be distinguished using structural mass spectrometry.

Andrey Dyachenko1, Ranit Gruber, Liat Shimon

  • 1Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.

Proceedings of the National Academy of Sciences of the United States of America
|April 17, 2013
PubMed
Summary

Structural mass spectrometry (MS) now allows researchers to determine all ligand binding constants for complex protein systems. This breakthrough enables the inference of allosteric mechanisms, advancing our understanding of protein regulation.

More Related Videos

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry
07:33

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry

Published on: October 15, 2018

Analyzing Large Protein Complexes by Structural Mass Spectrometry
15:35

Analyzing Large Protein Complexes by Structural Mass Spectrometry

Published on: June 19, 2010

Related Experiment Videos

Last Updated: May 12, 2026

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
10:01

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

Published on: November 28, 2017

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry
07:33

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry

Published on: October 15, 2018

Analyzing Large Protein Complexes by Structural Mass Spectrometry
15:35

Analyzing Large Protein Complexes by Structural Mass Spectrometry

Published on: June 19, 2010

Area of Science:

  • Biochemistry
  • Structural Biology
  • Biophysics

Background:

  • Protein activity is regulated by ligand-induced conformational changes, particularly in multimeric proteins.
  • Classic models (Monod-Wyman-Changeux, Koshland-Némethy-Filmer) describe allosteric cooperativity but are difficult to distinguish using bulk measurements.
  • Distinguishing allosteric mechanisms is crucial for understanding protein function and regulation.

Purpose of the Study:

  • To demonstrate how structural mass spectrometry (MS) can resolve ligand-bound states in multimeric proteins.
  • To enable determination of all binding constants in cooperative systems to infer allosteric mechanisms.
  • To provide a method applicable to diverse cooperative protein systems.

Main Methods:

  • Utilized advances in structural mass spectrometry (MS) to obtain the full distribution of ligand-bound states.
  • Developed novel data analysis techniques to interpret MS data for cooperative systems.
  • Validated the approach using the well-characterized Escherichia coli chaperone GroEL, a 14-site ATP-binding protein complex.

Main Results:

  • Successfully determined all 14 ATP binding constants for the Escherichia coli chaperone GroEL.
  • Characterized the ATP-loading pathway of GroEL, providing insights into its molecular machine function.
  • Demonstrated that structural MS can overcome limitations of ensemble measurements for dissecting allosteric mechanisms.

Conclusions:

  • Structural MS provides a powerful method to resolve ligand-bound states and determine binding constants in complex cooperative systems.
  • The methodology allows for the inference of allosteric mechanisms, previously challenging with bulk measurements.
  • This approach is broadly applicable to numerous cooperative systems, promising to accelerate research in protein allostery and molecular machines.