Structural stability of purified human CFTR is systematically improved by mutations in nucleotide binding domain 1.

ABSTRACT: The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is an ABC transporter containing two transmembrane domains forming a chloride ion channel, and two nucleotide binding domains (NBD1 and NBD2). CFTR has presented a formidable challenge to obtain monodisperse, biophysically stable protein. Here we report a comprehensive study comparing effects of single and multiple NBD1 mutations on stability of both the NBD1 domain alone and on purified full length human CFTR. Single mutations S492P, A534P, I539T acted additively, and when combined with M470V, S495P, and R555K cumulatively yielded an NBD1 with highly improved structural stability. Strategic combinations of these mutations strongly stabilized the domain to attain a calorimetric T_m > 70 °C. Replica exchange molecular

SUBMITTER: Yang Z 

PROVIDER: S-EPMC6319260 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature