TEMPO-Assisted Free-Radical-Initiated Peptide Sequencing Mass Spectrometry for Ubiquitin Ions: An Insight on the Gas-Phase Conformations

Lee, Jae-Ung; Lee, Sang Tak; Park, Chae Ri; Moon, Bongjin; Kim, Hugh I.; Oh, Han Bin

doi:10.1021/jasms.1c00313

Cited by 8 publications

(13 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The automatic gain control (AGC) was set to 2.0 x 10 -5 and the RF lens was set at 70%. The maximum injection time was set to 3000 and 5000 ms for MS 2 and MS 3 experiment, respectively. For all modified species, fifteen microscans were averaged and an isolation window of 5 thomsons was used.…”

Section: Mass Spectrometrymentioning

confidence: 99%

“…Free radical initiated peptide sequencing (FRIPS) is a radical-driven fragmentation technique that has been extensively applied to peptide ions. [1][2][3][4][5] Its major advantages over the predominantly utilized electron-driven technology (electron transfer dissociation, ETD) is its applicability to all peptides regardless of charge state, high duty cycle, and that it can be implemented on any mass spectrometer capable of collisional activation. 1,[3][4][5][6][7][8][9][10][11][12][13] These attributes stem from how the radical is initiated in each technique.…”

mentioning

confidence: 99%

“…Mobile proton-driven processes are often the lowest energy even-electron processes and have been avoided through the study of anions, sequestration of protons via the guanidation of lysine residues, development of a new FRIPS precursors, or focus on ions with fewer protons than arginine residues. 2,12,[16][17][18][19][20][21] Recently, a para-substituted FRIPS precursor was shown to more efficiently couple to peptides but its influence on the minimization of this undesired charge-directed processes has not been studied. 22 To be employed in more complex systems and proteomics workflows, the efficiency of sequence ion generation must be improved for multiply charged ions which are frequently generated in proteomics workflows.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Improved Performance of Positive-ion mode Free Radical-Initiated Peptide Sequencing with para-TEMPO-Bz

Osho

Wasik

Geary

et al. 2022

Preprint

View full text Add to dashboard Cite

Free radical-initiated peptide sequencing (FRIPS) is a tandem mass spectrometry (MS/MS) technique that generates sequence informative ions via collisionally-initiated radical chemistry. Collision activation (CA) homolytically cleaves an installed radical precursor, initiates radical formation, extensive hydrogen atom transfer, and peptide backbone dissociation. While FRIPS technique shows great promise, when applied to multiply charged derivatized peptide ions, a series of high abundance mass losses are observed which syphon ion abundance from radically generated sequence ions. This loss of ion abundance reduces the sequence coverage generated by FRIPS fragmentation. In this work, we hypothesized that these mass losses were instigated by the ortho-orientation of the radical precursor undergoing facile conversion into five- or six-membered intermediates and that the para-precursor would not undergo this chemistry. To test this assertion, we synthesized para-TEMPO-Bz, conjugated it to these peptides, and collisionally activated each. And indeed, we see the complete elimination of these undesired collisional processes and the significant increase in radical precursor ion abundance. The increase in ion abundance leads to a significant increase in the sequence coverage generated. These results demonstrate that p-TEMPO-Bz significantly improves the performance of positive-ion mode FRIPS and may be a suitable alternative to the currently utilized ortho-TEMPO-Bz-based FRIPS.

show abstract

Section: Mass Spectrometrymentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Improved Performance of Positive-ion mode Free Radical-Initiated Peptide Sequencing with para-TEMPO-Bz

Osho

Wasik

Geary

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Free radical initiated peptide sequencing (FRIPS) is a radical-driven fragmentation technique that has been extensively applied to peptide ions. [1][2][3][4][5] Its major advantages over the predominantly utilized electron-driven technology (electron transfer dissociation, ETD), is its applicability to all peptides, regardless of charge state, and that it can be implemented on any mass spectrometer capable of collisional activation. These attributes stem from how the radical is initiated in each technique.…”

mentioning

confidence: 99%

“…Mobile proton-driven processes are often the lowest energy even-electron processes and have been avoided through the study of anions, sequestration of protons via the guanidation of lysine residues, or focus on ions with fewer protons than arginine residues. 2,7,[9][10][11] To be employed in more complex systems and proteomics workflows, the efficiency of sequence ion generation must be improved for multiply charged ions which are frequently generated in proteomics workflows.…”

mentioning

confidence: 99%

Improved Performance of Positive-ion mode Free Radical-Initiated Peptide Sequencing with para-TEMPO-Bz

Osho

Wasik

Geary

et al. 2022

Preprint

View full text Add to dashboard Cite

Tandem mass spectrometry (MS/MS) is often employed for the sequencing of protein ions and the annotation of protein post-translational modification. Collision-induced dissociation (CID) and electron transfer dissociation (ETD) are the most frequently utilized MS/MS techniques, but both have significant limitations. CID is often funneled into a few kinet-ically labile bonds limiting the effectiveness of sequencing and ETD, is only applicable to multiply charged ions and re-quires specialized instrumentation. Free radical-initiated peptide sequencing (FRIPS) is an alternative fragmentation technique that generates sequence informative ions via collisionally-initiated radical chemistry. Its main advantages over the previously mentioned fragmentation techniques include charge state independence, improved efficiency of sequence ion generation, and applicability to most MS instrumentation. Collision activation (CA) homolytically cleaves the radical precursor, initiates radical formation, extensive hydrogen atom transfer, and peptide backbone dissociation. Although the FRIPS technique shows great promise when applied to cations, radical initiation competes directly with typical CID path-ways. When multiply charged ions of conjugated ACTH 1-14, b-amyloid 10-20, substance p, and melittin are collisionally activated, a series of high abundance mass losses are observed which syphon ion abundance from radically generated sequence ions. This loss of ion abundance reduces the sequence coverage generated by FRIPS fragmentation. In this work, we hypothesized that these mass losses were instigated by the ortho-orientation of the radical precursor undergoing fac-ile conversion into five- or six-membered intermediates and that the para-precursor would not undergo this chemistry. To test this assertion, we synthesized para-TEMPO-Bz, conjugated it to these peptides, and collisionally activated each. And indeed, we see the complete elimination of these undesired collisional processes and the significant increase in radi-cal precursor ion abundance. Moreover, we see a significant increase in the sequence coverage generated when the p-TEMPO-Bz is utilized. From these results, p-TEMPO-Bz significantly improves the performance of positive-ion mode FRIPS and may be a suitable alternative to the currently utilized ortho-TEMPO-Bz-based FRIPS.

show abstract

Thermodynamic Evaluation on Alkoxyamines of TEMPO Derivatives, Stable Alkoxyamines or Potential Radical Donors?

Qian

Zhang

et al. 2022

ChemistrySelect

View full text Add to dashboard Cite

In this work, the bond dissociation energy (BDE) values, including BDE(OÀ X) and BDE(NÀ O), for 129 alkoxyamines (XRT) of TEMPO derivatives were predicted, and the evaluations on stabilities of alkoxyamines, reactivities of alkoxyamines as radical donors and reactivity of TEMPO as various radical scavenger are well compared and discussed based on the predicted BDE values from the view of thermodynamic driving forces. Without a doubt, the wide and valuable BDE values are helpful for chemists better understand the thermodynamic properties and kinetic behaviors of alkoxyamines and TEMPO in chemical reactions and drug metabolism in vivo and promoting the rapid development and application of novel alkoxyamines.

show abstract

TEMPO-Assisted Free-Radical-Initiated Peptide Sequencing Mass Spectrometry for Ubiquitin Ions: An Insight on the Gas-Phase Conformations

Cited by 8 publications

References 64 publications

Improved Performance of Positive-ion mode Free Radical-Initiated Peptide Sequencing with para-TEMPO-Bz

Improved Performance of Positive-ion mode Free Radical-Initiated Peptide Sequencing with para-TEMPO-Bz

Improved Performance of Positive-ion mode Free Radical-Initiated Peptide Sequencing with para-TEMPO-Bz

Thermodynamic Evaluation on Alkoxyamines of TEMPO Derivatives, Stable Alkoxyamines or Potential Radical Donors?

Contact Info

Product

Resources

About