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 journalArticleResearchpeer-review

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

Fingerprint

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

@article{2a253dfd58eb47e6849ca20e883eb244,
title = "Development of Novel Free Radical Initiated Peptide Sequencing Reagent: Application to Identification and Characterization of Peptides by Mass Spectrometry",
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.].",
keywords = "Charge localize, Free radical, Hydrophobic peptides, Insulin, Peptide sequencing, Peptides without basic amino acid residues",
author = "Kaylee Gaspar and Kimberly Fabijanczuk and Tara Otegui and Jose Acosta and Jinshan Gao",
year = "2019",
month = "3",
day = "15",
doi = "10.1007/s13361-018-2114-8",
language = "English",
volume = "30",
pages = "548--556",
journal = "Journal of the American Society for Mass Spectrometry",
issn = "1044-0305",
publisher = "Springer New York",
number = "3",

}

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 journalArticleResearchpeer-review

TY - JOUR

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

PY - 2019/3/15

Y1 - 2019/3/15

N2 - 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.].

AB - 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.].

KW - Charge localize

KW - Free radical

KW - Hydrophobic peptides

KW - Insulin

KW - Peptide sequencing

KW - Peptides without basic amino acid residues

UR - http://www.scopus.com/inward/record.url?scp=85062838353&partnerID=8YFLogxK

U2 - 10.1007/s13361-018-2114-8

DO - 10.1007/s13361-018-2114-8

M3 - Article

VL - 30

SP - 548

EP - 556

JO - Journal of the American Society for Mass Spectrometry

JF - Journal of the American Society for Mass Spectrometry

SN - 1044-0305

IS - 3

ER -