title: Microwave-enhanced enzyme reaction for protein mapping by mass spectrometry: a new approach to protein digestion in minutes.
abstract: 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.
authors.affiliation: Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA. birendra.pramanik@spcorp.com
authors.forename: Birendra N
authors.initials: BN
authors.lastname: Pramanik
authors.forename: Urooj A
authors.initials: UA
authors.lastname: Mirza
authors.forename: Yao Hain
authors.initials: YH
authors.lastname: Ing
authors.forename: Yan-Hui
authors.initials: YH
authors.lastname: Liu
authors.forename: Peter L
authors.initials: PL
authors.lastname: Bartner
authors.forename: Patricia C
authors.initials: PC
authors.lastname: Weber
authors.forename: Ajay K
authors.initials: AK
authors.lastname: Bose
mesh_terms: 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
chemical_list: D003574:Cytochrome c Group; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D010446:Peptide Fragments; D011994:Recombinant Proteins; D025801:Ubiquitin; D014357:Trypsin