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

doi:https://doi.org/10.1021/ac010974p

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

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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 »« less

Authors:

Borisov, Oleg V ^[1]; Goshe, Michael B ^[2]; Conrads, Thomas P ^[2]; Rakov, Vsevolod S ^[2]; Veenstra, Timothy D ^[2]; Smith, Richard D ^[2]

ASSOC WESTERN UNIVERSITY
BATTELLE (PACIFIC NW LAB)

Publication Date:: 2002-05-15

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.

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                    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

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                    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.

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@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}

}

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