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Peptide Identification Using Tandem Mass Spectrometry

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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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Published on: July 31, 2011

Quantitative protein analysis by mass spectrometry.

Vishwajeeth R Pagala¹, Anthony A High, Xusheng Wang

¹St. Jude Proteomics Facility, St. Jude Children's Research Hospital, Memphis, TN, 38105, USA.

Methods in Molecular Biology (Clifton, N.J.)

|April 11, 2015

Summary

Mass spectrometry is a sensitive analytical technique now central to proteomics. This chapter details shotgun proteomics methods for protein identification, quantification, and post-translational modification analysis.

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Area of Science:

Analytical Chemistry
Proteomics

Background:

Mass spectrometry (MS) is a highly sensitive analytical chemistry method.
Its application in proteomics has grown significantly post-human genome sequencing.
MS is the primary technique for protein and post-translational modification identification and quantification.

Purpose of the Study:

To describe the experimental steps in a shotgun proteomics platform.
To cover sample preparation for protein-protein interaction studies.
To illustrate proteome quantification and bioinformatics analysis.

Main Methods:

Shotgun proteomics enables analysis of up to 10,000 proteins.
Detection sensitivity reaches the picogram range.
Key steps include sample preparation, MS data acquisition, and database searching.

Main Results:

Protein identification and post-translational modification analysis are detailed.
Proteome quantification strategies are presented.
Bioinformatics analysis is illustrated.

Conclusions:

Current shotgun proteomics platforms offer extensive capabilities.
Limitations and potential improvements for these platforms are discussed.