Development of Novel Free Radical Initiated Peptide Sequencing Reagent: Application to Identification and Characterization of Peptides by Mass Spectrometry

Kaylee Gaspar, Kimberly Fabijanczuk, Tara Otegui, Jose Acosta, Jinshan Gao

Research output: Contribution to journalArticle

Abstract

By incorporating a high proton affinity moiety to the charge localized free radical-initiated peptide sequencing (CL-FRIPS) reagent, FRIPS-MS technique has extended the applicability to hydrophobic peptides and peptides without basic amino acid residues (lysine, arginine, and histidine). Herein, the CL-FRIPS reagent has three moieties: (1) pyridine acting as the basic site to locate the proton, (2) 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO, a stable free radical) acting as the free radical precursor to generate the nascent free radical in the gas phase, and (3) N-hydroxysuccinimide (NHS) activated carboxylic acid acting as the coupling site to derivatize the N-terminus of peptides. The CL-FRIPS reagent allows for the characterization of peptides by generating sequencing ions, enzymatic cleavage-like radical-induced side chain losses, and the loss of TEMPO simultaneously via one-step collisional activation. Further collisional activation of enzymatic cleavage-like radical-induced side chain loss ions provides more information for the structure determination of peptides. The application of CL-FRIPS reagent to characterize peptides is proved by employing bovine insulin as the model peptide. Both scaffold structure of bovine insulin and sequencing information of each chain are achieved. [Figure not available: see fulltext.].

Original languageEnglish
Pages (from-to)548-556
Number of pages9
JournalJournal of the American Society for Mass Spectrometry
Volume30
Issue number3
DOIs
StatePublished - 15 Mar 2019

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Free Radicals
Mass spectrometry
Mass Spectrometry
Peptides
Protons
Chemical activation
Ions
Insulin
Basic Amino Acids
Carboxylic Acids
Histidine
Scaffolds
Lysine
Arginine
Gases

Keywords

  • Charge localize
  • Free radical
  • Hydrophobic peptides
  • Insulin
  • Peptide sequencing
  • Peptides without basic amino acid residues

Cite this

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abstract = "By incorporating a high proton affinity moiety to the charge localized free radical-initiated peptide sequencing (CL-FRIPS) reagent, FRIPS-MS technique has extended the applicability to hydrophobic peptides and peptides without basic amino acid residues (lysine, arginine, and histidine). Herein, the CL-FRIPS reagent has three moieties: (1) pyridine acting as the basic site to locate the proton, (2) 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO, a stable free radical) acting as the free radical precursor to generate the nascent free radical in the gas phase, and (3) N-hydroxysuccinimide (NHS) activated carboxylic acid acting as the coupling site to derivatize the N-terminus of peptides. The CL-FRIPS reagent allows for the characterization of peptides by generating sequencing ions, enzymatic cleavage-like radical-induced side chain losses, and the loss of TEMPO simultaneously via one-step collisional activation. Further collisional activation of enzymatic cleavage-like radical-induced side chain loss ions provides more information for the structure determination of peptides. The application of CL-FRIPS reagent to characterize peptides is proved by employing bovine insulin as the model peptide. Both scaffold structure of bovine insulin and sequencing information of each chain are achieved. [Figure not available: see fulltext.].",
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Development of Novel Free Radical Initiated Peptide Sequencing Reagent : Application to Identification and Characterization of Peptides by Mass Spectrometry. / Gaspar, Kaylee; Fabijanczuk, Kimberly; Otegui, Tara; Acosta, Jose; Gao, Jinshan.

In: Journal of the American Society for Mass Spectrometry, Vol. 30, No. 3, 15.03.2019, p. 548-556.

Research output: Contribution to journalArticle

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T1 - Development of Novel Free Radical Initiated Peptide Sequencing Reagent

T2 - Application to Identification and Characterization of Peptides by Mass Spectrometry

AU - Gaspar, Kaylee

AU - Fabijanczuk, Kimberly

AU - Otegui, Tara

AU - Acosta, Jose

AU - Gao, Jinshan

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KW - Peptide sequencing

KW - Peptides without basic amino acid residues

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