Premium
Identification of human myocardial proteins separated by two‐dimensional electrophoresis using an effective sample preparation for mass spectrometry
Author(s) -
Otto Albrecht,
Thiede Bernd,
Müller EvaChristina,
Scheler Christian,
WittmannLiebold Brigitte,
Jungblut Peter
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150171027
Subject(s) - chromatography , peptide mass fingerprinting , chemistry , gel electrophoresis , mass spectrometry , bottom up proteomics , sodium dodecyl sulfate , peptide , two dimensional gel electrophoresis , protein mass spectrometry , proteomics , biochemistry , tandem mass spectrometry , gene
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 improved 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.