Detection of Na_v1.5 Conformational Change in Mammalian Cells Using the Noncanonical Amino Acid ANAP.

ABSTRACT: Na_v1.5 inactivation is necessary for healthy conduction of the cardiac action potential. Genetic mutations of Na_v1.5 perturb inactivation and cause potentially fatal arrhythmias associated with long QT syndrome type 3. The exact structural dynamics of the inactivation complex is unknown. To sense inactivation gate conformational change in live mammalian cells, we incorporated the solvatochromic fluorescent noncanonical amino acid 3-((6-acetylnaphthalen-2-yl)amino)-2-aminopropanoic acid (ANAP) into single sites in the Na_v1.5 inactivation gate. ANAP was incorporated in full-length and C-terminally truncated Na_v1.5 channels using mammalian cell synthetase-tRNA technology. ANAP-incorporated channels were expressed in mammalian cells, and they exhibit

SUBMITTER: Shandell MA 

PROVIDER: S-EPMC6818161 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature