Targeted proteomics for metabolic pathway optimization

Tanveer S Batth, Jay D Keasling, Christopher J Petzold

20 Citations (Scopus)

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.
Original languageEnglish
Title of host publicationMethods in Molecular Biology
Number of pages13
Volume944
Publication date2012
Pages237-49
ISBN (Print)978-1-62703-121-9
ISBN (Electronic)978-1-62703-122-6
DOIs
Publication statusPublished - 2012
Externally publishedYes
SeriesMethods in molecular biology (Clifton, N.J.)

Keywords

  • Escherichia coli
  • Mass Spectrometry
  • Metabolic Networks and Pathways
  • Models, Molecular
  • Protein Conformation
  • Proteomics
  • Saccharomyces cerevisiae

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