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Protein identification and expression analysis using mass spectrometry.

Eugene Kolker1, Roger Higdon, Jason M Hogan

  • 1The BIATECH Institute, 19310 North Creek Parkway, Suite 115, Bothell, WA 98011, USA. ekolker@biatech.org

Trends in Microbiology
|April 11, 2006
PubMed
Summary
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This review details high-throughput proteomics, focusing on tandem mass spectrometry methods for reliable protein identification and quantification in laboratory settings. It covers experimental steps, challenges, and improvements for advancing proteomic analysis.

Area of Science:

  • Proteomics
  • Mass Spectrometry
  • Biotechnology

Background:

  • High-throughput proteomics aims to identify and quantify proteins expressed by organisms under specific conditions.
  • Advanced proteomic approaches yield novel biological data and hypotheses.
  • Understanding the capabilities and limitations of proteome studies in common labs is crucial.

Purpose of the Study:

  • To provide a practical report on the capabilities and limitations of proteome studies in laboratory settings.
  • To review popular tandem mass spectrometry-based methods for reliable proteomic analysis.
  • To discuss challenges and suggest improvements for proteomic workflows.

Main Methods:

  • Detailed step-by-step description of proteome experiments.
  • Focus on tandem mass spectrometry (MS/MS) techniques.

Related Experiment Videos

  • Includes sample preparation, digestion, labeling, liquid chromatography, data processing, database searching, and statistical analysis.

    • Main Results:

    • Discusses common difficulties and bottlenecks in proteome analysis.
    • Highlights requirements for enhancing the reliability and scope of proteomic studies.
    • Presents diverse high-throughput proteomics studies of microorganisms.

    Conclusions:

    • Tandem mass spectrometry is a key technique in high-throughput proteomics.
    • Reliable proteomic results require careful execution of experimental steps and data analysis.
    • Further improvements are needed to overcome current limitations in proteomic analysis.