TEMPO-Assisted Free Radical-Initiated Peptide Sequencing Mass Spectrometry (FRIPS MS) in Q-TOF and Orbitrap Mass Spectrometers: Single-Step Peptide Backbone Dissociations in Positive Ion Mode |
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| Authors: | Inae Jang Sun Young Lee Song Hwangbo Dukjin Kang Hookeun Lee Hugh I Kim Bongjin Moon Han Bin Oh |
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| Institution: | 1.Department of Chemistry,Sogang University,Seoul,Korea;2.College of Pharmacy,Kyung Hee University,Seoul,Korea;3.Center for Bioanalysis, Division of Metrology for Quality of Life,Korea Research Institute of Standards and Science,Daejeon,Korea;4.Gachon Institute of Pharmaceutical Sciences,Gachon University,Incheon,Korea;5.Department of Chemistry,Korea University,Seoul,Korea |
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| Abstract: | The present study demonstrates that one-step peptide backbone fragmentations can be achieved using the TEMPO 2-(2,2,6,6-tetramethyl piperidine-1-oxyl)]-assisted free radical-initiated peptide sequencing (FRIPS) mass spectrometry in a hybrid quadrupole time-of-flight (Q-TOF) mass spectrometer and a Q-Exactive Orbitrap instrument in positive ion mode, in contrast to two-step peptide fragmentation in an ion-trap mass spectrometer (reference Anal. Chem. 85, 7044–7051 (30)). In the hybrid Q-TOF and Q-Exactive instruments, higher collisional energies can be applied to the target peptides, compared with the low collisional energies applied by the ion-trap instrument. The higher energy deposition and the additional multiple collisions in the collision cell in both instruments appear to result in one-step peptide backbone dissociations in positive ion mode. This new finding clearly demonstrates that the TEMPO-assisted FRIPS approach is a very useful tool in peptide mass spectrometry research. |
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