Advertisement
MCP
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/mcp.M300045-MCP200 on June 26, 2003.
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
M300045-MCP200v1
2/7/463    most recent
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Glossary
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Janes, K. A.
Right arrow Articles by Yaffe, M. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Janes, K. A.
Right arrow Articles by Yaffe, M. B.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Molecular & Cellular Proteomics 2:463-473, 2003.
© 2003 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

A High-throughput Quantitative Multiplex Kinase Assay for Monitoring Information Flow in Signaling Networks

Application to Sepsis-Apoptosis*

Kevin A. Janes{ddagger}, John G. Albeck§, Lili X. Peng{ddagger}, Peter K. Sorger{ddagger},§,¶, Douglas A. Lauffenburger{ddagger},§,¶ and Michael B. Yaffe{ddagger},§,¶,||

From the {ddagger} Biological Engineering Division, § Department of Biology, and Center for Cancer Research, Massachusetts Institute of Technology, 400 Main Street, Cambridge, MA 02139

To treat complex human diseases effectively, a systems-level approach is needed to understand the interplay of environmental cues, intracellular signals, and cellular behaviors that underlie disease states. This approach requires high-throughput, multiplex techniques that measure quantitative temporal variations of multiple protein activities in the intracellular signaling network. Here, we describe a single microtiter-based format that simultaneously quantifies protein kinase activities in the phosphatidylinositol 3-kinase pathway (Akt), nuclear factor-{kappa}B pathway (IKK), and three core mitogen-activated protein kinase pathways (ERK, JNK1, MK2). These parallel high-throughput assays are stringently linear, redundantly specific, reproducible, and sensitive compared with classical low-throughput techniques. When applied to a model of sepsis-induced colon epithelial apoptosis, this approach identified a late phase of Akt activity as a critical mediator of cell survival that quantitatively contributed to the efficacy of insulin as an anti-apoptotic cue. Thus, sampling parallel nodes in the intracellular signaling network identified part of the molecular mechanism underlying the efficacy of insulin in the treatment of human sepsis.

|| To whom correspondence should be addressed: Center for Cancer Research, Massachusetts Institute of Technology, 77 Massachusetts Ave. E18-580, Cambridge, MA 02139. Tel.: 617-452-2103, Fax: 617-452-4978; E-mail: myaffe{at}mit.edu


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 Proc. Natl. Acad. Sci. USAHome page
Y. Yu, R. Anjum, K. Kubota, J. Rush, J. Villen, and S. P. Gygi
A site-specific, multiplexed kinase activity assay using stable-isotope dilution and high-resolution mass spectrometry
PNAS, July 14, 2009; 106(28): 11606 - 11611.
[Abstract] [Full Text] [PDF]

Home page
 Sci SignalHome page
R. C. Hsueh, M. Natarajan, I. Fraser, B. Pond, J. Liu, S. Mumby, H. Han, L. I. Jiang, M. I. Simon, R. Taussig, et al.
Deciphering Signaling Outcomes from a System of Complex Networks
Sci. Signal., May 19, 2009; 2(71): ra22 - ra22.
[Abstract] [Full Text] [PDF]

Home page
 Sci SignalHome page
A. Abdi, M. B. Tahoori, and E. S. Emamian
Fault Diagnosis Engineering of Digital Circuits Can Identify Vulnerable Molecules in Complex Cellular Pathways
Sci. Signal., October 21, 2008; 1(42): ra10 - ra10.
[Abstract] [Full Text] [PDF]

Home page
 Brief Funct Genomic ProteomicHome page
C. Jorgensen and R. Linding
Directional and quantitative phosphorylation networks
Briefings in Functional Genomics, February 12, 2008; (2008) eln001v1.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
J. E. McDunn, J. T. Muenzer, L. Rachdi, K. C. Chang, C. G. Davis, W. M. Dunne, D. Piwnica-Worms, E. Bernal-Mizrachi, and R. S. Hotchkiss
Peptide-mediated activation of Akt and extracellular regulated kinase signaling prevents lymphocyte apoptosis
FASEB J, February 1, 2008; 22(2): 561 - 568.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
E. J. Joslin, L. K. Opresko, A. Wells, H. S. Wiley, and D. A. Lauffenburger
EGF-receptor-mediated mammary epithelial cell migration is driven by sustained ERK signaling from autocrine stimulation
J. Cell Sci., October 15, 2007; 120(20): 3688 - 3699.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
M. L. Kemp, L. Wille, C. L. Lewis, L. B. Nicholson, and D. A. Lauffenburger
Quantitative Network Signal Combinations Downstream of TCR Activation Can Predict IL-2 Production Response
J. Immunol., April 15, 2007; 178(8): 4984 - 4992.
[Abstract] [Full Text] [PDF]

Home page
 Toxicol SciHome page
J. N. McDougal, C. M. Garrett, C. M. Amato, and S. J. Berberich
Effects of Brief Cutaneous JP-8 Jet Fuel Exposures on Time Course of Gene Expression in the Epidermis
Toxicol. Sci., February 1, 2007; 95(2): 495 - 510.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
K. Liby, M. Rendi, N. Suh, D. B. Royce, R. Risingsong, C. R. Williams, W. Lamph, F. Labrie, S. Krajewski, X. Xu, et al.
The Combination of the Rexinoid, LG100268, and a Selective Estrogen Receptor Modulator, Either Arzoxifene or Acolbifene, Synergizes in the Prevention and Treatment of Mammary Tumors in an Estrogen Receptor-Negative Model of Breast Cancer.
Clin. Cancer Res., October 1, 2006; 12(19): 5902 - 5909.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
K. Miller-Jensen, K. A. Janes, Y.-L. Wong, L. G. Griffith, and D. A. Lauffenburger
Adenoviral vector saturates Akt pro-survival signaling and blocks insulin-mediated rescue of tumor-necrosis-factor-induced apoptosis
J. Cell Sci., September 15, 2006; 119(18): 3788 - 3798.
[Abstract] [Full Text] [PDF]

Home page
 ScienceHome page
K. A. Janes, J. G. Albeck, S. Gaudet, P. K. Sorger, D. A. Lauffenburger, and M. B. Yaffe
A Systems Model of Signaling Identifies a Molecular Basis Set for Cytokine-Induced Apoptosis
Science, December 9, 2005; 310(5754): 1646 - 1653.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
S. Gaudet, K. A. Janes, J. G. Albeck, E. A. Pace, D. A. Lauffenburger, and P. K. Sorger
A Compendium of Signals and Responses Triggered by Prodeath and Prosurvival Cytokines
Mol. Cell. Proteomics, October 1, 2005; 4(10): 1569 - 1590.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
J.-E. Kim and S. R. Tannenbaum
Insulin Regulates Cleavage of Procaspase-9 via Binding of X Chromosome-Linked Inhibitor of Apoptosis Protein in HT-29 Cells
Cancer Res., December 15, 2004; 64(24): 9070 - 9075.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Journal of Biological Chemistry 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement