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

Graham, Nicholas A. and Pope, Melissa D. and Rimchala, Tharathorn and Huang, Beijing K. and Asthagiri, Anand R. (2007) A Microtiter Assay for Quantifying Protein-Protein Interactions Associated with Cell-Cell Adhesion. Journal of Biomolecular Screening, 12 (5). pp. 683-693. ISSN 1087-0571. doi:10.1177/1087057107301941. https://resolver.caltech.edu/CaltechAUTHORS:20170408-144719741

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Abstract

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 β-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 β-catenin bound to E-cadherin in unpurified whole-cell lysates with a Z′ factor of 0.74. The quantitative nature of the E-cadherin:β-catenin ELISA represents a dramatic improvement over the low-throughput assays currently used to characterize endogenous E-cadherin:β-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 β-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: β-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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1177/1087057107301941DOIArticle
Additional Information:© 2007 Society for Laboratory Automation and Screening. Received Nov 28, 2006, and in revised form Mar 2, 2007. Accepted for publication Mar 5, 2007. We thank Joan Brugge and David Schaffer for providing reagents. This work was supported by grants from the Concern Foundation for Cancer Research and the Jacobs Institute for Molecular Engineering for Medicine. NAG and MDP were partially supported by National Defense Science and Engineering and National Science Foundation graduate fellowships, respectively.
Funders:
Funding AgencyGrant Number
Concern Foundation for Cancer ResearchUNSPECIFIED
Caltech Jacobs Institute for Molecular Engineering for MedicineUNSPECIFIED
National Defense Science and Engineering Graduate (NDSEG) FellowshipUNSPECIFIED
NSF Graduate Research FellowshipUNSPECIFIED
Subject Keywords:E-cadherin, β-catenin, cell-cell adhesion, cancer, ELISA
Issue or Number:5
DOI:10.1177/1087057107301941
Record Number:CaltechAUTHORS:20170408-144719741
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20170408-144719741
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:76008
Collection:CaltechAUTHORS
Deposited By: 1Science Import
Deposited On:04 May 2017 21:09
Last Modified:15 Nov 2021 16:56

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