A Microtiter Assay for Quantifying Protein-Protein Interactions Associated with Cell-Cell Adhesion
| Autor: | Tharathorn Rimchala, Nicholas A. Graham, Beijing K. Huang, Anand R. Asthagiri, Melissa D. Pope |
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| Rok vydání: | 2007 |
| Předmět: |
0301 basic medicine
Cell Breast Neoplasms Enzyme-Linked Immunosorbent Assay Kidney Transfection medicine.disease_cause 01 natural sciences Biochemistry Cell Line Analytical Chemistry Protein–protein interaction Adherens junction Mice 03 medical and health sciences Cell Line Tumor Cell Adhesion medicine Animals Humans Cell adhesion beta Catenin biology Cadherin Antibodies Monoclonal Proteins Reproducibility of Results Epithelial Cells Cadherins Transmembrane protein 0104 chemical sciences Cell biology 010404 medicinal & biomolecular chemistry Retroviridae 030104 developmental biology medicine.anatomical_structure biology.protein Molecular Medicine Biological Assay Female Antibody Carcinogenesis Plasmids Biotechnology |
| Zdroj: | SLAS Discovery. 12:683-693 |
| ISSN: | 2472-5552 |
| DOI: | 10.1177/1087057107301941 |
| Popis: | 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 beta-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 beta-catenin bound to E-cadherin in unpurified whole-cell lysates with a Z' factor of 0.74. The quantitative nature of the E-cadherin:beta-catenin ELISA represents a dramatic improvement over the low-throughput assays currently used to characterize endogenous E-cadherin:beta-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 beta-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: beta-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. |
| Databáze: | OpenAIRE |
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