Biochemical characterization of the full-length isoform of soluble adenylyl cyclase
Year: 2025
Type: article
Abstract: cAMP is a ubiquitous second messenger produced from ATP and involved in many cellular processes. In humans, cAMP is produced from two types of adenylyl cyclases: G protein–regulated transmembrane adenylyl cyclases ( ADCY1-9 ) and bicarbonate- (HCO 3 − ) and calcium- (Ca 2+ ) regulated soluble adenylyl cyclase (sAC; ADCY10 ). sAC is molecularly and biochemically distinct from other mammalian nucleotidyl cyclases. In most mammals, sAC arises from a single gene which is predicted to generate multiple isoforms via alternative splicing. In rodents, there are two molecularly identified splice variants: the "full-length" isoform (sAC fl ) and a "truncated" isoform (sAC t ). To date, biochemical and structural characterization of sAC has focused almost exclusively on the sAC t isoform. Longer sAC isoforms, including the longest known sAC fl , contain additional presumptive regulatory domains which have not yet been functionally characterized. Thus far, studies have been limited by the inability to obtain sufficient sAC fl protein to allow in vitro biochemical characterization. Here, we describe attempts to heterologously express and purify human sAC fl as well as generation of a novel genetically modified mouse strain which permits biochemical separation and purification of endogenously expressed mouse sAC fl and sAC t . We use these heterologously expressed and endogenous proteins to compare and contrast the biochemically properties of human and mouse sAC fl and sAC t .
Source: Journal of Biological Chemistry
Institution: Weill Cornell Medicine
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,430
OpenAlex ID: W4415896175