Characterization of Adenylyl Cyclase Isoform 6 Residues Interacting with Forskolin.

ABSTRACT:

Background

The adenylyl cyclase (AC) pathway, crucial for pulmonary vasodilation, is inhibited by hypoxia. Forskolin (FSK) binds allosterically to AC, stimulating ATP catalysis. As AC6 is the primary AC isoform in the pulmonary artery, selective reactivation of AC6 could provide targeted reinstatement of hypoxic AC activity. This requires elucidation of the FSK binding site in AC6.

Methods

HEK293T cells stably overexpressing AC 5, 6, or 7 were incubated in normoxia (21% O₂) or hypoxia (10% O₂) or exposed to s-nitrosocysteine (CSNO). AC activity was measured using terbium norfloxacin assay; AC6 structure built by homology modeling; ligand docking to examine FSK-interacting amino acids; roles of selected residues determined by site-directed mutagenesis; F

SUBMITTER: Bhatia V 

PROVIDER: S-EPMC10135528 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature