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

Item Type:Article
Title:Identification of human myocardial proteins separated by two-dimensonal electrophoresis using an effective sample preparation for mass spectrometry
Creators Name:Otto, A. and Thiede, B. and Mueller, E.C. and Scheler, C. and Wittmann-Liebold, B. and Jungblut, P.
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.
Keywords:Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry, Peptide Mass Mapping, Two-Dimensional Gel Electrophoresis, Enzymatic In-Gel Digestion, Myocardial Proteins
Source:Electrophoresis
ISSN:0173-0835
Publisher:Wiley (Germany)
Volume:17
Number:10
Page Range:1643-1650
Date:1 October 1996
Official Publication:https://doi.org/10.1002/elps.1150171027
PubMed:View item in PubMed

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