New gating states of Cystic Fibrosis transmembrane conductance regulator discovered via studying pathogenic mutations, pharmacological reagents and ATP analogs.

Abstract

[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] Cystic Fibrosis (CF), caused by the loss of function in the gene for the protein cystic fibrosis transmembrane conductance (CFTR), is one of the most common lethal genetic diseases in the United States. By studying the structural/functional properties of CFTR, we are able to understand the molecular nature of this protein as well as to provide the potential target for drug designs. In my PhD study, I applied patch-clamp technique to explore the gating function of CFTR. Specifically, in my thesis, we approached a more complete CFTR gating scheme through three different ways. First, we proposed two new states need to be added to the original gating scheme to address unexpected results observed in G551D-CFTR, the third most common disease-associated mutant. Second, by studying the gating mechanism of NPPB on CFTR, we further confirmed the existence of these two states. Third, an ATP analog, 2-deoxy-ATP was used to explore the gating pathway provided by these two new states. This study not only bears the significance of the fundamental function of a single molecule, but also provides a new direction in the future drug development.