Journal of Biomolecular Screening

 

Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here to sign up for SAGE Journal Email Alerts today!

Sign In to gain access to subscriptions and/or personal tools.
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
1087057107301941v1
12/5/683    most recent
Right arrow References
Right arrow Alert me when this article is cited
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 ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Graham, N. A.
Right arrow Articles by Asthagiri, A. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Graham, N. A.
Right arrow Articles by Asthagiri, A. R.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?
This version was published on August 1, 2007
Journal of Biomolecular Screening, Vol. 12, No. 5, 683-693 (2007)
DOI: 10.1177/1087057107301941

A Microtiter Assay for Quantifying Protein-Protein Interactions Associated with Cell-Cell Adhesion

Nicholas A. Graham

Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena

Melissa D. Pope

Division of Engineering and Applied Science, California Institute of Technology, Pasadena

Tharathorn Rimchala

Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge

Beijing K. Huang

Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena

Anand R. Asthagiri

Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, anand{at}cheme.caltech.edu

Cell-cell adhesions are a hallmark of epithelial tissues, and the disruption of these contacts plays a critical role in both the early and late stages of oncogenesis. The interaction between the transmembrane protein E-cadherin and the intracellular protein ß-catenin plays a crucial role in the formation and maintenance of epithelial cell-cell contacts and is known to be downregulated in many cancers. The authors have developed a protein complex enzyme-linked immunosorbent assay (ELISA) that can quantify the amount of ß-catenin bound to E-cadherin in unpurified whole-cell lysates with a Z' factor of 0.74. The quantitative nature of the E-cadherin:ß-catenin ELISA represents a dramatic improvement over the low-throughput assays currently used to characterize endogenous E-cadherin:ß-catenin complexes. In addition, the protein complex ELISA format is compatible with standard sandwich ELISAs for parallel measurements of total levels of endogenous E-cadherin and ß-catenin. In 2 case studies closely related to cancer cell biology, the authors use the protein complex ELISA and traditional sandwich ELISAs to provide a detailed, quantitative picture of the molecular changes occurring within adherens junctions in vivo. Because the E-cadherin: ß-catenin protein complex plays a crucial role in oncogenesis, this protein complex ELISA may prove to be a valuable quantitative prognostic marker of tumor progression. (Journal of Biomolecular Screening 2007:683-693)

Key Words: E-cadherin • ß-catenin • cell-cell adhesion • cancer • ELISA


Add to CiteULike CiteULike   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?