Fluorescent isotope-coded affinity tag (FCAT) I

Design and synthesis

Zuly Rivera-Monroy, Guenther K. Bonn, A. Guttman

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A novel class of isotope-coded affinity tag is proposed possessing a fluorescent feature, referred to as fluorescent isotope-coded affinity tag (FCAT), to provide a new tool for quantitative proteomics. The label is designed to bind cysteine containing proteins or peptides. The FCAT reagent comprises four functional elements: a specific chemical reactivity group toward sulfhydryl groups; a linker that can incorporate the stable isotopes; a hydroxymethylbenzoic residue (base labile group) to cleave off a large part of the label before MS analysis; and a fluorescent tag for absolute quantification. The fluorescent part of the tag is also planned to be utilized to isolate the FCAT-labeled peptides via antibody based pull-down method. In this paper, we report on the solid phase organic synthesis of the light isotope containing FCAT molecule. The new labeling reagent showed good reactivity with model cysteine containing peptides. The fluorophore group was also effectively cleaved off from the labeled products to accommodate easier MS based analysis.

Original languageEnglish
Pages (from-to)299-311
Number of pages13
JournalBioorganic Chemistry
Volume36
Issue number6
DOIs
Publication statusPublished - Dec 2008

Fingerprint

Isotopes
Peptides
Cysteine
Labels
Synthetic Chemistry Techniques
Chemical reactivity
Solid-Phase Synthesis Techniques
Fluorophores
Proteomics
Labeling
Molecules
Antibodies
Proteins

Keywords

  • Isotope-coded reagents
  • Labeled peptides
  • MS/MS analysis
  • Proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry
  • Drug Discovery

Cite this

Fluorescent isotope-coded affinity tag (FCAT) I : Design and synthesis. / Rivera-Monroy, Zuly; Bonn, Guenther K.; Guttman, A.

In: Bioorganic Chemistry, Vol. 36, No. 6, 12.2008, p. 299-311.

Research output: Contribution to journalArticle

Rivera-Monroy, Zuly ; Bonn, Guenther K. ; Guttman, A. / Fluorescent isotope-coded affinity tag (FCAT) I : Design and synthesis. In: Bioorganic Chemistry. 2008 ; Vol. 36, No. 6. pp. 299-311.
@article{a86ebb18e94b4a3fbd43621796b0ddcb,
title = "Fluorescent isotope-coded affinity tag (FCAT) I: Design and synthesis",
abstract = "A novel class of isotope-coded affinity tag is proposed possessing a fluorescent feature, referred to as fluorescent isotope-coded affinity tag (FCAT), to provide a new tool for quantitative proteomics. The label is designed to bind cysteine containing proteins or peptides. The FCAT reagent comprises four functional elements: a specific chemical reactivity group toward sulfhydryl groups; a linker that can incorporate the stable isotopes; a hydroxymethylbenzoic residue (base labile group) to cleave off a large part of the label before MS analysis; and a fluorescent tag for absolute quantification. The fluorescent part of the tag is also planned to be utilized to isolate the FCAT-labeled peptides via antibody based pull-down method. In this paper, we report on the solid phase organic synthesis of the light isotope containing FCAT molecule. The new labeling reagent showed good reactivity with model cysteine containing peptides. The fluorophore group was also effectively cleaved off from the labeled products to accommodate easier MS based analysis.",
keywords = "Isotope-coded reagents, Labeled peptides, MS/MS analysis, Proteomics",
author = "Zuly Rivera-Monroy and Bonn, {Guenther K.} and A. Guttman",
year = "2008",
month = "12",
doi = "10.1016/j.bioorg.2008.08.005",
language = "English",
volume = "36",
pages = "299--311",
journal = "Bioorganic Chemistry",
issn = "0045-2068",
publisher = "Academic Press Inc.",
number = "6",

}

TY - JOUR

T1 - Fluorescent isotope-coded affinity tag (FCAT) I

T2 - Design and synthesis

AU - Rivera-Monroy, Zuly

AU - Bonn, Guenther K.

AU - Guttman, A.

PY - 2008/12

Y1 - 2008/12

N2 - A novel class of isotope-coded affinity tag is proposed possessing a fluorescent feature, referred to as fluorescent isotope-coded affinity tag (FCAT), to provide a new tool for quantitative proteomics. The label is designed to bind cysteine containing proteins or peptides. The FCAT reagent comprises four functional elements: a specific chemical reactivity group toward sulfhydryl groups; a linker that can incorporate the stable isotopes; a hydroxymethylbenzoic residue (base labile group) to cleave off a large part of the label before MS analysis; and a fluorescent tag for absolute quantification. The fluorescent part of the tag is also planned to be utilized to isolate the FCAT-labeled peptides via antibody based pull-down method. In this paper, we report on the solid phase organic synthesis of the light isotope containing FCAT molecule. The new labeling reagent showed good reactivity with model cysteine containing peptides. The fluorophore group was also effectively cleaved off from the labeled products to accommodate easier MS based analysis.

AB - A novel class of isotope-coded affinity tag is proposed possessing a fluorescent feature, referred to as fluorescent isotope-coded affinity tag (FCAT), to provide a new tool for quantitative proteomics. The label is designed to bind cysteine containing proteins or peptides. The FCAT reagent comprises four functional elements: a specific chemical reactivity group toward sulfhydryl groups; a linker that can incorporate the stable isotopes; a hydroxymethylbenzoic residue (base labile group) to cleave off a large part of the label before MS analysis; and a fluorescent tag for absolute quantification. The fluorescent part of the tag is also planned to be utilized to isolate the FCAT-labeled peptides via antibody based pull-down method. In this paper, we report on the solid phase organic synthesis of the light isotope containing FCAT molecule. The new labeling reagent showed good reactivity with model cysteine containing peptides. The fluorophore group was also effectively cleaved off from the labeled products to accommodate easier MS based analysis.

KW - Isotope-coded reagents

KW - Labeled peptides

KW - MS/MS analysis

KW - Proteomics

UR - http://www.scopus.com/inward/record.url?scp=54249154545&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=54249154545&partnerID=8YFLogxK

U2 - 10.1016/j.bioorg.2008.08.005

DO - 10.1016/j.bioorg.2008.08.005

M3 - Article

VL - 36

SP - 299

EP - 311

JO - Bioorganic Chemistry

JF - Bioorganic Chemistry

SN - 0045-2068

IS - 6

ER -