A tagging-via-substrate technology for detection and proteomics of farnesylated proteins

Yoonjung Kho, Sung Chan Kim, Chen Jiang, Deb Barma, Sung Won Kwon, Jinke Cheng, Janis Jaunbergs, Carolyn Weinbaum, Fuyuhiko Tamanoi, J R Falck, Yingming Zhao

Research output: Contribution to journalArticle

247 Citations (Scopus)

Abstract

A recently developed proteomics strategy, designated tagging-via-substrate (TAS) approach, is described for the detection and proteomic analysis of farnesylated proteins. TAS technology involves metabolic incorporation of a synthetic azido-farnesyl analog and chemoselective derivatization of azido-farnesyl-modified proteins by an elegant version of Staudinger reaction, pioneered by the Bertozzi group, using a biotinylated phosphine capture reagent. The resulting protein conjugates can be specifically detected and/or affinity-purified by streptavidin-linked horseradish peroxidase or agarose beads, respectively. Thus, the technology enables global profiling of farnesylated proteins by enriching farnesylated proteins and reducing the complexity of farnesylation subproteome. Azido-farnesylated proteins maintain the properties of protein farnesylation, including promoting membrane association, Ras-dependent mitogen-activated protein kinase kinase activation, and inhibition of lovastatin-induced apoptosis. A proteomic analysis of farnesylated proteins by TAS technology revealed 18 farnesylated proteins, including those with potentially novel farnesylation motifs, suggesting that future use of this method is likely to yield novel insight into protein farnesylation. TAS technology can be extended to other posttranslational modifications, such as geranylgeranylation and myristoylation, thus providing powerful tools for detection, quantification, and proteomic analysis of posttranslationally modified proteins.

Original languageEnglish (US)
Pages (from-to)12479-12484
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number34
DOIs
StatePublished - Aug 24 2004

Fingerprint

Proteomics
Technology
Prenylation
Proteins
Protein Prenylation
phosphine
Lovastatin
Streptavidin
Mitogen-Activated Protein Kinase Kinases
Horseradish Peroxidase
Post Translational Protein Processing
Sepharose
Apoptosis
Membranes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

A tagging-via-substrate technology for detection and proteomics of farnesylated proteins. / Kho, Yoonjung; Kim, Sung Chan; Jiang, Chen; Barma, Deb; Kwon, Sung Won; Cheng, Jinke; Jaunbergs, Janis; Weinbaum, Carolyn; Tamanoi, Fuyuhiko; Falck, J R; Zhao, Yingming.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 101, No. 34, 24.08.2004, p. 12479-12484.

Research output: Contribution to journalArticle

Kho, Y, Kim, SC, Jiang, C, Barma, D, Kwon, SW, Cheng, J, Jaunbergs, J, Weinbaum, C, Tamanoi, F, Falck, JR & Zhao, Y 2004, 'A tagging-via-substrate technology for detection and proteomics of farnesylated proteins', Proceedings of the National Academy of Sciences of the United States of America, vol. 101, no. 34, pp. 12479-12484. https://doi.org/10.1073/pnas.0403413101
Kho, Yoonjung ; Kim, Sung Chan ; Jiang, Chen ; Barma, Deb ; Kwon, Sung Won ; Cheng, Jinke ; Jaunbergs, Janis ; Weinbaum, Carolyn ; Tamanoi, Fuyuhiko ; Falck, J R ; Zhao, Yingming. / A tagging-via-substrate technology for detection and proteomics of farnesylated proteins. In: Proceedings of the National Academy of Sciences of the United States of America. 2004 ; Vol. 101, No. 34. pp. 12479-12484.
@article{f1c739934e844c10bc449f02e75a7176,
title = "A tagging-via-substrate technology for detection and proteomics of farnesylated proteins",
abstract = "A recently developed proteomics strategy, designated tagging-via-substrate (TAS) approach, is described for the detection and proteomic analysis of farnesylated proteins. TAS technology involves metabolic incorporation of a synthetic azido-farnesyl analog and chemoselective derivatization of azido-farnesyl-modified proteins by an elegant version of Staudinger reaction, pioneered by the Bertozzi group, using a biotinylated phosphine capture reagent. The resulting protein conjugates can be specifically detected and/or affinity-purified by streptavidin-linked horseradish peroxidase or agarose beads, respectively. Thus, the technology enables global profiling of farnesylated proteins by enriching farnesylated proteins and reducing the complexity of farnesylation subproteome. Azido-farnesylated proteins maintain the properties of protein farnesylation, including promoting membrane association, Ras-dependent mitogen-activated protein kinase kinase activation, and inhibition of lovastatin-induced apoptosis. A proteomic analysis of farnesylated proteins by TAS technology revealed 18 farnesylated proteins, including those with potentially novel farnesylation motifs, suggesting that future use of this method is likely to yield novel insight into protein farnesylation. TAS technology can be extended to other posttranslational modifications, such as geranylgeranylation and myristoylation, thus providing powerful tools for detection, quantification, and proteomic analysis of posttranslationally modified proteins.",
author = "Yoonjung Kho and Kim, {Sung Chan} and Chen Jiang and Deb Barma and Kwon, {Sung Won} and Jinke Cheng and Janis Jaunbergs and Carolyn Weinbaum and Fuyuhiko Tamanoi and Falck, {J R} and Yingming Zhao",
year = "2004",
month = "8",
day = "24",
doi = "10.1073/pnas.0403413101",
language = "English (US)",
volume = "101",
pages = "12479--12484",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "34",

}

TY - JOUR

T1 - A tagging-via-substrate technology for detection and proteomics of farnesylated proteins

AU - Kho, Yoonjung

AU - Kim, Sung Chan

AU - Jiang, Chen

AU - Barma, Deb

AU - Kwon, Sung Won

AU - Cheng, Jinke

AU - Jaunbergs, Janis

AU - Weinbaum, Carolyn

AU - Tamanoi, Fuyuhiko

AU - Falck, J R

AU - Zhao, Yingming

PY - 2004/8/24

Y1 - 2004/8/24

N2 - A recently developed proteomics strategy, designated tagging-via-substrate (TAS) approach, is described for the detection and proteomic analysis of farnesylated proteins. TAS technology involves metabolic incorporation of a synthetic azido-farnesyl analog and chemoselective derivatization of azido-farnesyl-modified proteins by an elegant version of Staudinger reaction, pioneered by the Bertozzi group, using a biotinylated phosphine capture reagent. The resulting protein conjugates can be specifically detected and/or affinity-purified by streptavidin-linked horseradish peroxidase or agarose beads, respectively. Thus, the technology enables global profiling of farnesylated proteins by enriching farnesylated proteins and reducing the complexity of farnesylation subproteome. Azido-farnesylated proteins maintain the properties of protein farnesylation, including promoting membrane association, Ras-dependent mitogen-activated protein kinase kinase activation, and inhibition of lovastatin-induced apoptosis. A proteomic analysis of farnesylated proteins by TAS technology revealed 18 farnesylated proteins, including those with potentially novel farnesylation motifs, suggesting that future use of this method is likely to yield novel insight into protein farnesylation. TAS technology can be extended to other posttranslational modifications, such as geranylgeranylation and myristoylation, thus providing powerful tools for detection, quantification, and proteomic analysis of posttranslationally modified proteins.

AB - A recently developed proteomics strategy, designated tagging-via-substrate (TAS) approach, is described for the detection and proteomic analysis of farnesylated proteins. TAS technology involves metabolic incorporation of a synthetic azido-farnesyl analog and chemoselective derivatization of azido-farnesyl-modified proteins by an elegant version of Staudinger reaction, pioneered by the Bertozzi group, using a biotinylated phosphine capture reagent. The resulting protein conjugates can be specifically detected and/or affinity-purified by streptavidin-linked horseradish peroxidase or agarose beads, respectively. Thus, the technology enables global profiling of farnesylated proteins by enriching farnesylated proteins and reducing the complexity of farnesylation subproteome. Azido-farnesylated proteins maintain the properties of protein farnesylation, including promoting membrane association, Ras-dependent mitogen-activated protein kinase kinase activation, and inhibition of lovastatin-induced apoptosis. A proteomic analysis of farnesylated proteins by TAS technology revealed 18 farnesylated proteins, including those with potentially novel farnesylation motifs, suggesting that future use of this method is likely to yield novel insight into protein farnesylation. TAS technology can be extended to other posttranslational modifications, such as geranylgeranylation and myristoylation, thus providing powerful tools for detection, quantification, and proteomic analysis of posttranslationally modified proteins.

UR - http://www.scopus.com/inward/record.url?scp=4344584507&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4344584507&partnerID=8YFLogxK

U2 - 10.1073/pnas.0403413101

DO - 10.1073/pnas.0403413101

M3 - Article

C2 - 15308774

AN - SCOPUS:4344584507

VL - 101

SP - 12479

EP - 12484

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 34

ER -