Analysis of phospho- and glycopolypeptides with infrared matrix-assisted laser desorption and ionization.
The analytical characteristics of infrared (IR) matrix-assisted laser desorption and ionization (MALDI) were investigated for the analysis of phosphopeptides, a phosphopolypeptide, and glycopeptides. Two commercially available instruments, a high-resolution delayed extraction (DE) reflectron time-of-flight (RETOF) mass spectrometer and a high-power pulsed Er:YAG laser, were interfaced to produce a high-resolution MALDI-DE-RETOF instrument that is easy to use and can be switched between UV- and IR-MALDI mode within seconds. In the interface design, particular attention was paid to maintaining the same professional operating environment for the new IR-MALDI mode as exists for the commercial UV-MALDI mode. This instrument configuration facilitates comparative observation and investigation of the relative analytical merits of IR- and UV-MALDI. The results of studies of the tryptic alpha-casein phosphopeptides, RP1 (a Thr45-monophosphorylated congener of the recombinant protein hirudin variant 1), and fetuin Asn81 tryptic glycopeptides are presented. The elimination of labile substituents such as phosphoric acid and sialic acid is suppressed in IR-MALDI-RETOF mass spectrometry, with concomitant higher analyte ion yields. These results reflect the advantages that accrue from deposition of significantly less internal energy in the case of IR-MALDI.
Cramer
R
R
Ludwig Institute for Cancer Research, London, U.K.
Richter
W J
WJ
Stimson
E
E
Burlingame
A L
AL
D000595:Amino Acid Sequence; D002364:Caseins / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006020:Glycopeptides / Q000032:analysis*; D006629:Hirudins / Q000737:chemistry; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods; D010748:Phosphopeptides / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013055:Spectrophotometry, Infrared; D013056:Spectrophotometry, Ultraviolet; D014357:Trypsin / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000737:chemistry
D002364:Caseins; D006020:Glycopeptides; D006629:Hirudins; D010748:Phosphopeptides; D000509:alpha-Fetoproteins; D014357:Trypsin