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Title: Low-Energy Collision-Induced Dissociation Fragmentation Analysis of Cysteinyl-Modified Peptides

Abstract

The development of methods to chemically modify and isolate cysteinyl-residue containing peptides (Cys-peptides) for LC-MS/MS analysis has generated considerable interest in the field of proteomics. Methods using isotope-coded affinity tags (ICAT) and (+)-biotinyl-iodoacetamidyl-3,6-dioxaoctanediamine (iodoacetyl-PEO-biotin) employ similar Cys-modifying reagents that contain a thiolate-specific biotin group to modify and isolate Cys-containing peptides in conjunction with immobilized avidin. For these strategies to be effective on a proteome-wide level, the presence of the ICAT or acetyl-PEO-biotin tag should not interfere with the efficiency of induced dissociation in MS/MS experiments or with the identification of the modified Cys-peptides by automated database searching algorithms. We have compared the collision-induced dissociation (CID) fragmentation patterns of peptides labeled with iodoacetyl-PEO-biotin and the ICAT reagent to those of the unmodified peptides. CID of Cys-peptides modified with either reagent resulted in the formation of ions attributed to the modified Cys-peptides as well as those unique to the labeling reagent. As demonstrated by analyzing acetyl-PEO-biotin labeled peptides from ribonuclease A and the ICAT-labeled proteome of D. radiodurans, the presence of these labeled-specific product ions provides a useful identifier to discern whether a peptide has been modified with the Cys-specific reagent, especially when a number of peptides analyzed using these methods domore » not contain a modified Cys-residue, and to differentiate identical Cys-peptides labeled with either ICAT-D0 or ICAT-D8.« less

Authors:
 [1];  [2];  [2];  [2];  [2];  [2]
  1. ASSOC WESTERN UNIVERSITY
  2. BATTELLE (PACIFIC NW LAB)
Publication Date:
Research Org.:
Pacific Northwest National Lab., Richland, WA (US)
Sponsoring Org.:
US Department of Energy (US)
OSTI Identifier:
15002842
Report Number(s):
PNWD-SA-5627
TRN: US200420%%89
DOE Contract Number:  
AC06-76RL01830
Resource Type:
Journal Article
Journal Name:
Analytical Chemistry
Additional Journal Information:
Journal Volume: 74; Other Information: PBD: 15 May 2002
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; AFFINITY; ALGORITHMS; AVIDIN; BIOTIN; DISSOCIATION; EFFICIENCY; FRAGMENTATION; PEPTIDES; RNA-ASE

Citation Formats

Borisov, Oleg V, Goshe, Michael B, Conrads, Thomas P, Rakov, Vsevolod S, Veenstra, Timothy D, and Smith, Richard D. Low-Energy Collision-Induced Dissociation Fragmentation Analysis of Cysteinyl-Modified Peptides. United States: N. p., 2002. Web. doi:10.1021/ac010974p.
Borisov, Oleg V, Goshe, Michael B, Conrads, Thomas P, Rakov, Vsevolod S, Veenstra, Timothy D, & Smith, Richard D. Low-Energy Collision-Induced Dissociation Fragmentation Analysis of Cysteinyl-Modified Peptides. United States. https://doi.org/10.1021/ac010974p
Borisov, Oleg V, Goshe, Michael B, Conrads, Thomas P, Rakov, Vsevolod S, Veenstra, Timothy D, and Smith, Richard D. Wed . "Low-Energy Collision-Induced Dissociation Fragmentation Analysis of Cysteinyl-Modified Peptides". United States. https://doi.org/10.1021/ac010974p.
@article{osti_15002842,
title = {Low-Energy Collision-Induced Dissociation Fragmentation Analysis of Cysteinyl-Modified Peptides},
author = {Borisov, Oleg V and Goshe, Michael B and Conrads, Thomas P and Rakov, Vsevolod S and Veenstra, Timothy D and Smith, Richard D},
abstractNote = {The development of methods to chemically modify and isolate cysteinyl-residue containing peptides (Cys-peptides) for LC-MS/MS analysis has generated considerable interest in the field of proteomics. Methods using isotope-coded affinity tags (ICAT) and (+)-biotinyl-iodoacetamidyl-3,6-dioxaoctanediamine (iodoacetyl-PEO-biotin) employ similar Cys-modifying reagents that contain a thiolate-specific biotin group to modify and isolate Cys-containing peptides in conjunction with immobilized avidin. For these strategies to be effective on a proteome-wide level, the presence of the ICAT or acetyl-PEO-biotin tag should not interfere with the efficiency of induced dissociation in MS/MS experiments or with the identification of the modified Cys-peptides by automated database searching algorithms. We have compared the collision-induced dissociation (CID) fragmentation patterns of peptides labeled with iodoacetyl-PEO-biotin and the ICAT reagent to those of the unmodified peptides. CID of Cys-peptides modified with either reagent resulted in the formation of ions attributed to the modified Cys-peptides as well as those unique to the labeling reagent. As demonstrated by analyzing acetyl-PEO-biotin labeled peptides from ribonuclease A and the ICAT-labeled proteome of D. radiodurans, the presence of these labeled-specific product ions provides a useful identifier to discern whether a peptide has been modified with the Cys-specific reagent, especially when a number of peptides analyzed using these methods do not contain a modified Cys-residue, and to differentiate identical Cys-peptides labeled with either ICAT-D0 or ICAT-D8.},
doi = {10.1021/ac010974p},
url = {https://www.osti.gov/biblio/15002842}, journal = {Analytical Chemistry},
number = ,
volume = 74,
place = {United States},
year = {2002},
month = {5}
}