Improved infrared multiphoton dissociation of peptides through N-terminal phosphonite derivatization |
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Authors: | Lisa A Vasicek Jeffrey J Wilson Jennifer S Brodbelt |
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Institution: | (1) Department of Chemistry and Biochemistry, The University of Texas at Austin, 1 University Station A5300, Austin, TX 78712, USA;(2) Departments of Chemistry and Biomolecular Chemistry, University of Wisconsin, Madison, WI 53706, USA;(3) Thermo Fisher Scientific GmbH, Bremen, Germany; |
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Abstract: | A strategy for improving the sequencing of peptides by infrared multiphoton dissociation (IRMPD) in a linear ion trap mass
spectrometer is described. We have developed an N-terminal derivatization reagent, 4-methylphosphonophenylisothiocyanate (PPITC),
which allows the attachment of an IR-chromogenic phosphonite group to the N-terminus of peptides, thus enhancing their IRMPD
efficiencies. After the facile derivatization process, the PPITC-modified peptides require shorter irradiation times for efficient
IRMPD and yield extensive series of y ions, including those of low m/z that are not detected upon traditional CID. The resulting IRMPD mass spectra afford more complete sequence coverage for both
model peptides and tryptic peptides from cytochrome c. We compare the effectiveness of this derivatization/IRMPD approach to that of a common N-terminal sulfonation reaction that
utilizes 4-sulfophenylisothiocyanate (SPITC) in conjunction with CID and IRMPD. |
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