A new application of microwave technology to proteomics.
Two-dimensional electrophoresis (2-DE) combined with mass spectrometry has significantly improved the possibilities of large-scale identification of proteins. However, 2-DE is limited by its inability to speed up the in-gel digestion process. We have developed a new approach to speed up the protein identification process utilizing microwave technology. Proteins excised from gels are subjected to in-gel digestion with endoprotease trypsin by microwave irradiation, which rapidly produces peptide fragments. The peptide fragments were further analyzed by matrix-assisted laser desorption/ionization technique for protein identification. The efficacy of this technique for protein mapping was demonstrated by the mass spectral analyses of the peptide fragmentation of several proteins, including lysozyme, albumin, conalbumin, and ribonuclease A. The method reduced the required time for in-gel digestion of proteins from 16 hours to as little as five minutes. This new application of microwave technology to protein identification will be an important advancement in biotechnology and proteome research.
Juan
Hsueh-Fen
HF
Institute of Molecular and Cellular Biology, National Taiwan University, Taipei. yukijuan@ntu.edu.tw
Chang
Shing-Chuan
SC
Huang
Hsuan-Cheng
HC
Chen
Shui-Tein
ST
D000418:Albumins / Q000737:chemistry; D000818:Animals; D002417:Cattle; D002645:Chickens; D003207:Conalbumin / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D008872:Microwaves; D009113:Muramidase / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods; D010455:Peptides / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000418:Albumins; D010455:Peptides; D020543:Proteome; D003207:Conalbumin; D012259:Ribonuclease, Pancreatic; D009113:Muramidase