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Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

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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...
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Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

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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...
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Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

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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 electrospray 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...
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High-Resolution Mass Spectrometry (HRMS)01:15

High-Resolution Mass Spectrometry (HRMS)

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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...
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Mass Spectrum: Interpretation01:24

Mass Spectrum: Interpretation

1.1K
An unknown compound can be established by identifying the molecular ion peak in the mass spectrum. The molecular ion peak is often weak or absent due to the predominance of fragmentation in high-energy electron beams. In such cases, a low-energy electron beam can be used to scan the spectrum to enhance the intensity of the molecular ion peak. Additionally, chemical ionization, field ionization, and desorption ionization spectra are used to obtain a relatively intense molecular ion peak.
To...
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Mass Analyzers: Overview01:13

Mass Analyzers: Overview

599
The mass analyzer is a crucial component of the mass spectrometer. In the ionization chamber, the vaporized sample is bombarded with a high-energy electron beam to generate a radical cation and further fragment into neutral molecules, radicals, and cations. A series of negatively charged accelerator plates accelerate the cations into the mass analyzer. The mass analyzer separates ions according to their mass-to-charge (m/z) ratios and then directs them to the detector. The common types of mass...
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Related Experiment Video

Updated: Jun 9, 2025

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

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A Conformation-Specific Approach to Native Top-down Mass Spectrometry.

Hannah M Britt1, Aisha Ben-Younis1, Nathanael Page1,2

  • 1Institute of Structural and Molecular Biology, University College London, London WC1E 6BT, United Kingdom.

Journal of the American Society for Mass Spectrometry
|October 25, 2024
PubMed
Summary

This study introduces a new native top-down mass spectrometry method using ion mobility to analyze protein conformations. This technique reveals conformation-specific structural insights and proteoform variations, even in protein-ligand complexes.

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Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry
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Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry

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

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Last Updated: Jun 9, 2025

Analyzing Protein Architectures and Protein-Ligand Complexes by Integrative Structural Mass Spectrometry
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Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry
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Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry

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

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Structural Biology

Background:

  • Native top-down mass spectrometry (MS) is valuable for proteoform characterization.
  • Current MS methods offer limited insights into protein conformation, often in bulk or without direct measurement.
  • There is a need for conformation-specific structural analysis in MS.

Purpose of the Study:

  • To develop a novel ion-mobility-enabled native top-down MS method.
  • To enable conformation-specific characterization of proteins and their complexes.
  • To provide direct structural insights linked to specific protein conformations.

Main Methods:

  • Implementation of ion mobility coupled with native top-down mass spectrometry.
  • Analysis of the model protein calmodulin to identify conformation-linked differences.
  • Application to protein-ligand complexes for component identification and structural probing.

Main Results:

  • Demonstrated conformation-specific differences in backbone dissociation for calmodulin.
  • Successfully linked proteoform variations with distinct structural conformations.
  • Showcased the method's utility in analyzing protein-ligand interactions and ligand-induced changes.

Conclusions:

  • The new method provides unprecedented conformation-specific structural information using native top-down MS.
  • This approach advances the characterization of proteoforms and protein dynamics.
  • The technique is versatile for studying both free proteins and their complexes.