High‐performance human myocardial two‐dimensional electrophoresis database: Edition 1996

Abstract

The master gel of the human myocardial two-dimensional electrophoresis (2-DE) gel database contains about 3300 protein spots characterized in terms of isoelectric point (pI) and molecular mass. A high-performance technique was applied, using large gels (23 x 30 cm). Isoelectric focusing with anodic sample preparation and nonequilibrium running conditions (NEPHGE) was combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in 15% acrylamide gels in the second dimension. The range of pI extends from pH 4.5 to 9.6. Seventy proteins were identified by combinations of amino acid analysis, N-terminal and internal sequencing, immunostaining, matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS) peptide mass fingerprinting, post-source decay MALDI-MS and ladder sequencing by carboxypeptidase P. The identification of additional proteins, not found in the master gel, was achieved by immunoblotting. Unequivocal identification with high sensitivity and good yield was obtained by combining internal sequencing and MALDI-MS. In-gel digestion, the concentration and purification of peptides in a peptide collecting device, and the improved FRAGMOD program for peptide mass fingerprinting have added to the security and sensitivity of identification. The high-performance human myocardial 2-DE database was built up with proteins detected by the TOPSPOT program. Spots within six sections of the whole pattern are clickable. Protein description includes detailed information about identification, characterization, and links to the related SWISS-PROT, other 2-DE databases and Medline entries. The database is constructed in accordance with four of the rules for a federated database.

Study of Burkitt lymphoma cell line proteins by high resolution two-dimensional gel electrophoresis and nanoelectrospray mass spectrometry

“…Triosephosphate isomerase (TPI), present in spot D29, catalyzes the interconversion of dihydroxy acetone phosphate and glyceraldehyde 3-phosphate in glycolysis, glucogenesis, fatty acid synthesis, and the pentose shunt. It is the product of a single gene; however, two electrophoretic forms were reported [7,13] which may result from deaminations at Asn-71 and Asn-15 [41]. We observed modification of cysteine and methionine, and the Asn-71-containing peptide was not deaminated; the Asn-15-containing peptide was not detected.…”

Proteome reference map and regulation network of neonatal rat cardiomyocyte

“…As mentioned earlier, the protein database will be important to serve as a basis for further studies of human disease. There are four 2-DE protein databases of cardiac proteins, established by three independent groups, which can be accessed via the World Wide Web [7,[14][15][16] . These databases facilitate proteomic research into heart diseases containing information In addition, 2-DE protein databases and proteomic maps for other mammals are also under construction to support work on animal models of heart disease [17][18][19][20] .…”

Proteomic analysis of whey from bovine colostrum and mature milk

“…Proteins were stained with colloidal Coomassie Brilliant Blue G-250. Two-dimensional gels were analyzed by Topspot 2.0 software (Algorithmus, Prehm, Berlin), previously used by Müller et al (1996).…”