Accelerated identification of proteins by mass spectrometry by employing covalent pre-gel staining with Uniblue A.

PloS One
Marco A Mata-GómezRobert Winkler

Abstract

The identification of proteins by mass spectrometry is a standard method in biopharmaceutical quality control and biochemical research. Prior to identification by mass spectrometry, proteins are usually pre-separated by electrophoresis. However, current protein staining and de-staining protocols are tedious and time consuming, and therefore prolong the sample preparation time for mass spectrometry. We developed a 1-minute covalent pre-gel staining protocol for proteins, which does not require de-staining before the mass spectrometry analysis. We investigated the electrophoretic properties of derivatized proteins and peptides and studied their behavior in mass spectrometry. Further, we elucidated the preferred reaction of proteins with Uniblue A and demonstrate the integration of the peptide derivatization into typical informatics tools. The Uniblue A staining method drastically speeds up the sample preparation for the mass spectrometry based identification of proteins. The application of this chemo-proteomic strategy will be advantageous for routine quality control of proteins and for time-critical tasks in protein analysis.

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Datasets Mentioned

BETA
DB00073

Methods Mentioned

BETA
electrophoresis
acetylation
glycosylations
chip

Software Mentioned

CompassXport
mmass
Open Mass Spectrometry Search Algorithm ( OMSSA
PRoteomics IDEntifications
pl
bat
FastaTools
OMSSAVALIDATION
PeptideProphet
VennDiagram

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