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Targeted Proteomics for Metabolic Pathway Optimization

  • Tanveer S. Batth
  • Jay D. Keasling
  • Christopher J. PetzoldEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 944)

Abstract

A crucial part of optimization of metabolically engineered organisms is producing balanced levels of pathway proteins. Typically, protein levels are monitored by Western blot analysis; however, application to multiple enzyme pathways can be difficult without unique antibodies for each enzyme in the pathway. Furthermore, it can be time consuming, and cost prohibitive during exploratory stages of pathway design when many different proteins must be monitored simultaneously. We present here a targeted proteomics approach that uses selected-reaction monitoring (SRM) mass spectrometry to quantify multiple proteins in a sample. SRM methods provide high selectivity and high sensitivity to enable rapid quantification of multiple proteins in an engineered pathway regardless of sequence or organism of origin.

Key words

Targeted proteomics Selected-reaction monitoring mass spectrometry Protein quantification Metabolic pathway optimization 

Notes

Acknowledgments

This work conducted by the Joint BioEnergy Institute was supported by the Office of Science, Office of Biological and Environmental Research, of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Tanveer S. Batth
    • 1
  • Jay D. Keasling
    • 2
    • 3
  • Christopher J. Petzold
    • 2
    Email author
  1. 1.Physical Biosciences DivisionLawrence Berkeley National Laboratory, Joint BioEnergy Institute (JBEI)BerkeleyUSA
  2. 2.Physical Biosciences DivisionJoint BioEnergy Institute (JBEI), Lawrence Berkeley National LaboratoryBerkeleyUSA
  3. 3.Department of Chemical & Biomolecular Engineering, Department of Bioengineering, Berkeley Center for Synthetic Biology (SynBERC)University of CaliforniaBerkeleyUSA

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