Identification of human myocardial proteins separated by two-dimensional electrophoresis using an effective sample preparation for mass spectrometry

Electrophoresis
A OttoP Jungblut

Abstract

Peptide mass fingerprinting is a powerful tool for the identification of proteins separated by two-dimensional gel electrophoresis (2-DE). The identification of in-gel digested proteins by peptide mass fingerprinting was significantly improve in comparison to blot-digests by using a peptide-collecting device. This device allows the effective purification and concentration of enzymatic digests of low-intensity spots without expensive equipment and is described in detail. Sensitivity in the fmol range was demonstrated by unequivocal identification of bovine serum albumin after sodium dodecyl sulfate--polyacrylamide gel electrophoresis. Furthermore the high performance liquid chromatography pattern of in-gel digests indicated a 2- to 3-fold higher yield of the separated peptides. Therefore, a higher amount of the peptides was available to perform N-terminal sequencing. The identification of 16 proteins from a high-resolution 2-DE gel map of human myocardium tissue has been achieved by means of this technique. Three of these proteins were associated with changes in spot intensity with dilated cardiomyopathy.

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Citations

May 1, 1997·Mass Spectrometry Reviews·P Jungblut, B Thiede
May 27, 1999·Electrophoresis·X P LiP R Jungblut
Jul 12, 2002·Proteomics·Chantragan SrisomsapJisnuson Svasti
Aug 1, 1997·Electrophoresis·I Humphery-SmithW P Blackstock
Sep 5, 2003·Journal of the American Society for Mass Spectrometry·William J HenzelJohn T Stults
Sep 5, 2003·Journal of the American Society for Mass Spectrometry·Frank SchmidtKlaus Peter Pleissner
Sep 1, 2001·Trends in Cardiovascular Medicine·J Macri, S T Rapundalo
Sep 1, 2001·Trends in Cardiovascular Medicine·J D Cavalcoli
Feb 1, 2000·Drug Discovery Today·M J Dunn
May 23, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·D JägerU Müller-Werdan
Dec 11, 2002·European Journal of Biochemistry·Matthias BrockWolfgang Buckel
Aug 21, 2003·Human Molecular Genetics·Emma McGregor, Michael J Dunn
May 11, 2002·Journal of Bacteriology·Tanja EpplerWinfried Boos
Jan 16, 2002·Journal of Bacteriology·Tamara HoffmannErhard Bremer
Feb 20, 2004·Journal of Bacteriology·Teresa CostaAdriano O Henriques
Mar 19, 2014·Journal of Proteomics·Manuel AlfaroAntonio G Pisabarro
Jul 23, 2003·Journal of Cellular Biochemistry·Qing-Yu He, Jen-Fu Chiu

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