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.
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. Graphical Abstract ᅟ.
Department of Chemistry, Sogang University, Seoul, 04107, Korea.
Inae
I
Jang
College of Pharmacy, Kyung Hee University, Seoul, 02447, Korea.
Sun Young
SY
Lee
Department of Chemistry, Sogang University, Seoul, 04107, Korea.
Song
S
Hwangbo
Center for Bioanalysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon, 34113, Korea.
Dukjin
D
Kang
Gachon Institute of Pharmaceutical Sciences, Gachon University, Incheon, 21936, Korea.
Hookeun
H
Lee
Department of Chemistry, Korea University, Seoul, 02841, Korea.
Hugh I
HI
Kim
Department of Chemistry, Sogang University, Seoul, 04107, Korea.
Bongjin
B
Moon
Department of Chemistry, Sogang University, Seoul, 04107, Korea. hanbinoh@sogang.ac.kr.
Han Bin
0000-0001-7919-0393
HB
Oh
D000595:Amino Acid Sequence; D000818:Animals; D001920:Bradykinin / Q000737:chemistry; D002417:Cattle; D003497:Cyclic N-Oxides / Q000737:chemistry*; D045304:Cytochromes c / Q000737:chemistry; D005609:Free Radicals / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D020539:Sequence Analysis, Protein / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D003497:Cyclic N-Oxides; D005609:Free Radicals; D010446:Peptide Fragments; D010455:Peptides; D045304:Cytochromes c; D001920:Bradykinin; C003959:TEMPO
Free radical-initiated peptide sequencing (FRIPS); Orbitrap; Q-TOF; Radical-based fragmentations; TEMPO
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