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Related Concept Videos

Electrospray Ionization (ESI) Mass Spectrometry01:12

Electrospray Ionization (ESI) Mass Spectrometry

Higher molecular weight biomolecules are nonvolatile compounds that may decompose before ionizing or vaporizing during mass analysis with conventional electron impact ionization methods. Accordingly, electrospray ionization (ESI) is the favored method for vaporizing and ionizing biomolecules as it circumvents rapid fragmentation and enables the recording of mass signals for the entire biomolecule.
ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the...
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...

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Analyzing Large Protein Complexes by Structural Mass Spectrometry
15:35

Analyzing Large Protein Complexes by Structural Mass Spectrometry

Published on: June 19, 2010

Increasing charge while preserving noncovalent protein complexes for ESI-MS.

Shirley H Lomeli1, Sheng Yin, Rachel R Ogorzalek Loo

  • 1Department of Chemistry and Biochemistry, University of California-Los Angeles, CA 90095, USA.

Journal of the American Society for Mass Spectrometry
|December 23, 2008
PubMed
Summary

Adding m-nitrobenzyl alcohol (m-NBA) to native protein solutions enhances multiple charging in electrospray ionization mass spectrometry. This "supercharging" effect increases protein complex detection and facilitates tandem mass spectrometry studies.

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Last Updated: Jun 26, 2026

Analyzing Large Protein Complexes by Structural Mass Spectrometry
15:35

Analyzing Large Protein Complexes by Structural Mass Spectrometry

Published on: June 19, 2010

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

Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Mass Spectrometry

Background:

  • Native protein analysis via mass spectrometry is crucial for understanding complex biological systems.
  • Electrospray ionization (ESI) is a key technique for analyzing intact proteins and complexes.
  • Optimizing charge states is essential for sensitive detection and structural studies.

Purpose of the Study:

  • To investigate the effect of m-nitrobenzyl alcohol (m-NBA) on the charging of native proteins and noncovalent complexes.
  • To assess the potential of m-NBA as a 'supercharging' agent in ESI mass spectrometry.
  • To evaluate the utility of enhanced charging for mass spectrometry-based protein complex analysis.

Main Methods:

  • Nondenaturing protein solutions were prepared with varying concentrations of m-nitrobenzyl alcohol (m-NBA).
  • Electrospray ionization (ESI) mass spectra were acquired for native proteins and protein complexes.
  • Charge state distributions were analyzed and compared to control samples without m-NBA.

Main Results:

  • Addition of m-NBA significantly increased the multiple charging of native proteins and noncovalent protein complexes.
  • Observed charge increases ranged from 8% for large proteasome complexes to 48% for smaller proteins.
  • No dissociation of noncovalently bound subunits was detected in the presence of m-NBA.

Conclusions:

  • m-Nitrobenzyl alcohol acts as an effective 'supercharging' agent in ESI mass spectrometry for native proteins.
  • Enhanced charging improves mass spectrometer detection limits and facilitates tandem mass spectrometry of protein complexes.
  • This method offers a valuable approach for studying large and weakly bound protein assemblies.