Profiling the activity of G proteins in patient-derived tissues by rapid affinity-capture of signal transduction proteins (GRASP)

David M. Berman, Ie Ming Shih, Lorri Anne Burke, Timothy D. Veenstra, Yingming Zhao, Thomas P. Conrads, Sung Won Kwon, Van Hoang, Li Rong Yu, Ming Zhou, Robert J. Kurman, Emanuel F. Petricoin, Lance A. Liotta

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The next phase in molecular medicine will require the ability to identify signal transduction events inside a cell, in the biologic context of the disease-host interface and at a given point in time. New technologies are needed to profile the activity of these signaling pathways in patient tissue rather than cultured cell lines since the tumor-host microenvironment influences the cellular proteome. We introduce such a technology, rapid affinity capture of signaling proteins (GRASP), to investigate the activity of signaling pathways from patient-derived carcinomas and benign epithelial surfaces and apply it to studying important signaling events in ovarian carcinoma. During the progression from benign ovarian epithelium to invasive carcinoma, there is loss of repression of Rho A as evidenced by its dissociation from its inhibitor, Rho Guanine Nucleotide Dissociation Inhibitor (RhoGDI). GRASP is more informative than simply profiling transcript or protein levels. Furthermore, GRASP coupled with mass spectrometry allowed us to identify a protein-binding partner of RhoGDI, demonstrating the power of this technology in the discovery of potentially novel protein-protein interactions. GRASP represents an advance in the field of proteomics as it detects protein interactions present in cells as they exist in their native tissue microenvironment.

Original languageEnglish (US)
Pages (from-to)812-818
Number of pages7
JournalProteomics
Volume4
Issue number3
DOIs
StatePublished - Mar 2004

Fingerprint

Signal transduction
GTP-Binding Proteins
Signal Transduction
rho-Specific Guanine Nucleotide Dissociation Inhibitors
Tissue
Proteins
Technology
Carcinoma
Molecular Medicine
Tumor Microenvironment
Proteome
Protein Binding
Proteomics
Medicine
Mass spectrometry
Tumors
Cultured Cells
Mass Spectrometry
Epithelium
Cells

Keywords

  • Affinity capture
  • G protein signal transduction
  • Mass spectrometry
  • Ovarian carcinoma
  • Protein-protein interactions

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Berman, D. M., Shih, I. M., Burke, L. A., Veenstra, T. D., Zhao, Y., Conrads, T. P., ... Liotta, L. A. (2004). Profiling the activity of G proteins in patient-derived tissues by rapid affinity-capture of signal transduction proteins (GRASP). Proteomics, 4(3), 812-818. https://doi.org/10.1002/pmic.200300579

Profiling the activity of G proteins in patient-derived tissues by rapid affinity-capture of signal transduction proteins (GRASP). / Berman, David M.; Shih, Ie Ming; Burke, Lorri Anne; Veenstra, Timothy D.; Zhao, Yingming; Conrads, Thomas P.; Kwon, Sung Won; Hoang, Van; Yu, Li Rong; Zhou, Ming; Kurman, Robert J.; Petricoin, Emanuel F.; Liotta, Lance A.

In: Proteomics, Vol. 4, No. 3, 03.2004, p. 812-818.

Research output: Contribution to journalArticle

Berman, DM, Shih, IM, Burke, LA, Veenstra, TD, Zhao, Y, Conrads, TP, Kwon, SW, Hoang, V, Yu, LR, Zhou, M, Kurman, RJ, Petricoin, EF & Liotta, LA 2004, 'Profiling the activity of G proteins in patient-derived tissues by rapid affinity-capture of signal transduction proteins (GRASP)', Proteomics, vol. 4, no. 3, pp. 812-818. https://doi.org/10.1002/pmic.200300579
Berman, David M. ; Shih, Ie Ming ; Burke, Lorri Anne ; Veenstra, Timothy D. ; Zhao, Yingming ; Conrads, Thomas P. ; Kwon, Sung Won ; Hoang, Van ; Yu, Li Rong ; Zhou, Ming ; Kurman, Robert J. ; Petricoin, Emanuel F. ; Liotta, Lance A. / Profiling the activity of G proteins in patient-derived tissues by rapid affinity-capture of signal transduction proteins (GRASP). In: Proteomics. 2004 ; Vol. 4, No. 3. pp. 812-818.
@article{6a8caf6b35eb4980817cb78bd9abddd8,
title = "Profiling the activity of G proteins in patient-derived tissues by rapid affinity-capture of signal transduction proteins (GRASP)",
abstract = "The next phase in molecular medicine will require the ability to identify signal transduction events inside a cell, in the biologic context of the disease-host interface and at a given point in time. New technologies are needed to profile the activity of these signaling pathways in patient tissue rather than cultured cell lines since the tumor-host microenvironment influences the cellular proteome. We introduce such a technology, rapid affinity capture of signaling proteins (GRASP), to investigate the activity of signaling pathways from patient-derived carcinomas and benign epithelial surfaces and apply it to studying important signaling events in ovarian carcinoma. During the progression from benign ovarian epithelium to invasive carcinoma, there is loss of repression of Rho A as evidenced by its dissociation from its inhibitor, Rho Guanine Nucleotide Dissociation Inhibitor (RhoGDI). GRASP is more informative than simply profiling transcript or protein levels. Furthermore, GRASP coupled with mass spectrometry allowed us to identify a protein-binding partner of RhoGDI, demonstrating the power of this technology in the discovery of potentially novel protein-protein interactions. GRASP represents an advance in the field of proteomics as it detects protein interactions present in cells as they exist in their native tissue microenvironment.",
keywords = "Affinity capture, G protein signal transduction, Mass spectrometry, Ovarian carcinoma, Protein-protein interactions",
author = "Berman, {David M.} and Shih, {Ie Ming} and Burke, {Lorri Anne} and Veenstra, {Timothy D.} and Yingming Zhao and Conrads, {Thomas P.} and Kwon, {Sung Won} and Van Hoang and Yu, {Li Rong} and Ming Zhou and Kurman, {Robert J.} and Petricoin, {Emanuel F.} and Liotta, {Lance A.}",
year = "2004",
month = "3",
doi = "10.1002/pmic.200300579",
language = "English (US)",
volume = "4",
pages = "812--818",
journal = "Proteomics",
issn = "1615-9853",
publisher = "Wiley-VCH Verlag",
number = "3",

}

TY - JOUR

T1 - Profiling the activity of G proteins in patient-derived tissues by rapid affinity-capture of signal transduction proteins (GRASP)

AU - Berman, David M.

AU - Shih, Ie Ming

AU - Burke, Lorri Anne

AU - Veenstra, Timothy D.

AU - Zhao, Yingming

AU - Conrads, Thomas P.

AU - Kwon, Sung Won

AU - Hoang, Van

AU - Yu, Li Rong

AU - Zhou, Ming

AU - Kurman, Robert J.

AU - Petricoin, Emanuel F.

AU - Liotta, Lance A.

PY - 2004/3

Y1 - 2004/3

N2 - The next phase in molecular medicine will require the ability to identify signal transduction events inside a cell, in the biologic context of the disease-host interface and at a given point in time. New technologies are needed to profile the activity of these signaling pathways in patient tissue rather than cultured cell lines since the tumor-host microenvironment influences the cellular proteome. We introduce such a technology, rapid affinity capture of signaling proteins (GRASP), to investigate the activity of signaling pathways from patient-derived carcinomas and benign epithelial surfaces and apply it to studying important signaling events in ovarian carcinoma. During the progression from benign ovarian epithelium to invasive carcinoma, there is loss of repression of Rho A as evidenced by its dissociation from its inhibitor, Rho Guanine Nucleotide Dissociation Inhibitor (RhoGDI). GRASP is more informative than simply profiling transcript or protein levels. Furthermore, GRASP coupled with mass spectrometry allowed us to identify a protein-binding partner of RhoGDI, demonstrating the power of this technology in the discovery of potentially novel protein-protein interactions. GRASP represents an advance in the field of proteomics as it detects protein interactions present in cells as they exist in their native tissue microenvironment.

AB - The next phase in molecular medicine will require the ability to identify signal transduction events inside a cell, in the biologic context of the disease-host interface and at a given point in time. New technologies are needed to profile the activity of these signaling pathways in patient tissue rather than cultured cell lines since the tumor-host microenvironment influences the cellular proteome. We introduce such a technology, rapid affinity capture of signaling proteins (GRASP), to investigate the activity of signaling pathways from patient-derived carcinomas and benign epithelial surfaces and apply it to studying important signaling events in ovarian carcinoma. During the progression from benign ovarian epithelium to invasive carcinoma, there is loss of repression of Rho A as evidenced by its dissociation from its inhibitor, Rho Guanine Nucleotide Dissociation Inhibitor (RhoGDI). GRASP is more informative than simply profiling transcript or protein levels. Furthermore, GRASP coupled with mass spectrometry allowed us to identify a protein-binding partner of RhoGDI, demonstrating the power of this technology in the discovery of potentially novel protein-protein interactions. GRASP represents an advance in the field of proteomics as it detects protein interactions present in cells as they exist in their native tissue microenvironment.

KW - Affinity capture

KW - G protein signal transduction

KW - Mass spectrometry

KW - Ovarian carcinoma

KW - Protein-protein interactions

UR - http://www.scopus.com/inward/record.url?scp=12144285847&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=12144285847&partnerID=8YFLogxK

U2 - 10.1002/pmic.200300579

DO - 10.1002/pmic.200300579

M3 - Article

VL - 4

SP - 812

EP - 818

JO - Proteomics

JF - Proteomics

SN - 1615-9853

IS - 3

ER -