Identification of proteins in cell-free extracts using liquid chromatography-electrospray ionization mass spectrometry

Joseph J. Dalluge, Prasad Reddy

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

A rapid method for the identification and characterization of proteins in bacterial cell-free extracts has been developed using directly combined liquid chromatography-electrospray mass spectrometry. The usefulness of this technique is demonstrated for monitoring the expression and chemical modification of phosphoenolpyruvate-sugar phosphotransferase system (PTS) proteins from E. coli with molecular masses ranging from 9-65 kDa. The technique is characterized by minimal sample preparation, remarkable mass accuracy and resolution, reproducibility and the ability, unlike gel electrophoresis, to directly identify post-translational modifications. The advantages of this technique over analogous matrix-assisted laser desorption ionization mass spectrometry approaches and its potential as a standard tool in the biomolecular research laboratory are discussed.

Original languageEnglish (US)
Pages (from-to)156-160
Number of pages5
JournalBioTechniques
Volume28
Issue number1
DOIs
StatePublished - 2000

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