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

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 1 2004

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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  • Cite this

    Berman, D. M., Shih, I. M., Burke, L. A., Veenstra, T. D., Zhao, Y., Conrads, T. P., Kwon, S. W., Hoang, V., Yu, L. R., Zhou, M., Kurman, R. J., Petricoin, E. F., & 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