Detecting Interaction of CFTR Polypeptides
Cystic fibrosis (CF) is the most common genetic disease of Caucasians in North America, occurring at a frequency of approximately 1 in 2500 births. The disease results from defective function of the gene encoding the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein in a variety of tissues, including the pancreas and the lung epithelium.
The present invention describes materials and methods for detecting the interaction of CFTR proteins. The method can be used to determine whether one CFTR NBD1 polypeptide interacts with a second CFTR NBD1 polypeptide using a yeast dual hybrid assay. The subject methods can be used to determine whether mutations to the CFTR polypeptide reduce or eliminate dimerization of the CFTR polypeptides. The present methods can also be used to screen and identify revertant mutations that restore dimerization of a mutant CFTR polypeptide, as well as mutations that enhance dimerization and CFTR activity greater than that of wildtype protein.The subject invention also provides materials and methods for efficiently identifying and screening for compounds, drugs and other such compositions that facilitate proper dimerization of the CFTR polypeptides and would be candidate agents for use in treating patients having CF.
- The use of a yeast growth bioassay is fast and inexpensive in comparison to current screening procedures that involve mammalian cells and assays for CFTR channel activity
- Can be used to evaluate a large number of compounds in a high throughput format