Friedman, Thomas (NIDCD)
Belyantseva, Inna (NIDCD)
Proof of concept has been demonstrated.
Investigators at the National Institute on Deafness and Other Communication Disorders (NIDCD) have developed an assay for the detection of protein-protein interactions in living cells. This assay uses readily-available reagents and straightforward techniques that avoid the difficulty of purifying proteins or generating antibodies required for other binding studies. Proof-of-concept for this assay has been demonstrated, and a manuscript is in preparation for publication.
This technology utilizes a molecular motor, myosin X, which migrates along actin filaments within cells. A protein fused to a fragment of myosin X will carry its binding partners to the cell periphery. Since the myosin fusion protein and its partner are labeled with different fluorescent tags, an unambiguous fluorescence overlap will be visible as discrete points along the periphery of the cell. The inventors have designed a number of cDNAs for the construction of fusion proteins appropriate for such an assay.
Available for licensing are a variety of cDNAs which may be used for generating fluorescently-tagged myosin X fusion proteins, for use in the assay described above. Also available are a number of constructs incorporating other fluorescently-tagged myosins, kinesins, myosin and kinesin binding partners and a variety of PDZ scaffold proteins. Further details of the available cDNAs are available upon request.
- Identification of protein-protein binding interactions in living cells
- DNA-based tools for study of myosins, trafficking, signaling complexes and other research focusing on molecular motors
- Assay avoids the need to purify proteins or generate antibodies for binding studies
- Protein-protein interactions can be unambiguously identified