Intact protein analysis by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry.
Direct tandem mass spectrometric (MS/MS) analysis of small, singly charged protein ions by tandem time-of-flight mass spectrometry (TOFMS) is demonstrated for proteins up to a molecular mass of 12 kDa. The MALDI-generated singly charged precursor ions predominantly yield product ions resulting from metastable fragmentation at aspartyl and prolyl residues. Additional series of C-terminal sequence ions provide in some cases sufficient information for protein identification. The amount of sample required to obtain good quality spectra is in the high femtomolar to low picomolar range. Within this range, MALDI-MS/MS using TOF/TOF trade mark ion optics now provides the opportunity for direct protein identification and partial characterization without prior enzymatic hydrolysis.
Lin
Melanie
M
Applied Biosystems, Framingham, MA 01701, USA. linmx@appliedbiosystems.com
Campbell
Jennifer M
JM
Mueller
Dieter R
DR
Wirth
Urs
U
D000818:Animals; D002417:Cattle; D006629:Hirudins / Q000032:analysis; D007328:Insulin / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D011994:Recombinant Proteins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation / Q000379:methods*; D013879:Thioredoxins / Q000032:analysis
D006629:Hirudins; D007328:Insulin; D011506:Proteins; D011994:Recombinant Proteins; D013879:Thioredoxins