Microwave-enhanced enzyme reaction for protein mapping by mass spectrometry: a new approach to protein digestion in minutes.
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 alpha-2b. Most important, using this novel approach digestion of proteins occurs in minutes, in contrast to the hours required by conventional methods.
Pramanik
Birendra N
BN
Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA. birendra.pramanik@spcorp.com
Mirza
Urooj A
UA
Ing
Yao Hain
YH
Liu
Yan-Hui
YH
Bartner
Peter L
PL
Weber
Patricia C
PC
Bose
Ajay K
AK
D000818:Animals; D002417:Cattle; D003574:Cytochrome c Group / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008872:Microwaves; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010449:Peptide Mapping / Q000379:methods*; D011994:Recombinant Proteins; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism
D003574:Cytochrome c Group; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D010446:Peptide Fragments; D011994:Recombinant Proteins; D025801:Ubiquitin; D014357:Trypsin