Microwave‐enhanced enzyme reaction for protein mapping by mass spectrometry: A new approach to protein digestion in minutes

• Published in: Protein science Vol. 11; no. 11; pp. 2676 - 2687
• Main Authors: Pramanik, Birendra N., Mirza, Urooj A., Ing, Yao Hain, Liu, Yan‐Hui, Bartner, Peter L., Weber, Patricia C., Bose, Ajay K.
• Format: Journal Article
• Language: English
• Published: Bristol Cold Spring Harbor Laboratory Press 01-11-2002
• Subjects: accelerated protein digestion; Animals; Cattle; Cytochrome c Group - chemistry; Cytochrome c Group - genetics; Cytochrome c Group - metabolism; Interferon-alpha - chemistry; Interferon-alpha - genetics; Interferon-alpha - metabolism; mass spectrometry; Microwave irradiation; Microwaves; Molecular Weight; Peptide Fragments - chemistry; Peptide Fragments - genetics; Peptide Fragments - metabolism; Peptide Mapping - methods; Recombinant Proteins; selective endoprotease reactions; Spectrometry, Mass, Electrospray Ionization; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Time Factors; Trypsin - metabolism; Ubiquitin - chemistry; Ubiquitin - metabolism
• ISSN: 0961-8368; 1469-896X
• DOI: 10.1110/ps.0213702

Accelerated proteolytic cleavage of proteins under controlled microwave irradiation has been achieved. Selective peptide fragmentation by endoproteases trypsin or lysine C led to smaller peptides that were analyzed by matrix‐assisted laser desorption ionization (MALDI) or liquid chromatography‐electrospray ionization (LC‐ESI) techniques. The efficacy of this technique for protein mapping was demonstrated by the mass spectral analyses of the peptide fragmentation of several biologically active proteins, including cytochrome c, ubiquitin, lysozyme, myoglobin, and interferon α‐2b. Most important, using this novel approach digestion of proteins occurs in minutes, in contrast to the hours required by conventional methods.