Determining the molecular interactions within CFTR that result in Cystic Fibrosis - Pilot Project of the COBRE for the Center for Molecular Medicine

Cystic Fibrosis (CF) is a recessive genetic disease resulting from any one of over 1000 unique mutations to the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR). These mutations disrupt the channel which ultimately results in loss of hydration across the lung epithelium preventing mucocilliary clearance, leading to colonization of bacteria, followed by destruction of the lung through chronic infection. 70% of CF patients are homozygous for the deletion of phenylalanine at position 508 (ÄF508) in CFTR. A quantitative description of the impact this deletion mutation makes on the stability of this five domain channel, will be determined. The mechanistic details of CFTR delivery will be measured following the fluorescence of SEP engineered into CFTR. The rate of delivery, number of CFTR channels deposited, and cell surface stability will be quantified for wild-type and ÄF508-CFTR in the presence of small molecule modulators. The role that assembly of CFTR with ENaC plays in the trafficking of CFTR will be quantified by FRET. Understanding how molecular defects effect stability and the role played in trafficking should allow for the design of improved therapeutics to reverse the molecular defect caused by a single mutation in the CFTR channel.