Molecular and biochemical characterization of the bicarbonate-sensing soluble adenylyl cyclase from a bony fish, the rainbow trout Oncorhynchus mykiss .

Autor: Salmerón C; Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, USA.; Department of Pharmacology, University of California San Diego, San Diego, CA, USA., Harter TS; Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, USA., Kwan GT; Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, USA., Roa JN; Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, USA., Blair SD; Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.; Department of Biology, Winthrop University, Rock Hill, SC, USA., Rummer JL; Australian Research Council Centre of Excellence for Coral Reef Studies, James Cook University, Townsville, Queensland, Australia., Shiels HA; Division of Cardiovascular Sciences, Faculty of Biology, Medicine and Health, University of Manchester, UK., Goss GG; Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada., Wilson RW; Department of Biosciences, University of Exeter, Exeter, UK., Tresguerres M; Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, USA.
Jazyk: angličtina
Zdroj: Interface focus [Interface Focus] 2021 Apr 06; Vol. 11 (2), pp. 20200026. Date of Electronic Publication: 2021 Feb 12.
DOI: 10.1098/rsfs.2020.0026
Abstrakt: Soluble adenylyl cyclase (sAC) is a HC O 3   - -stimulated enzyme that produces the ubiquitous signalling molecule cAMP, and deemed an evolutionarily conserved acid-base sensor. However, its presence is not yet confirmed in bony fishes, the most abundant and diverse of vertebrates. Here, we identified sAC genes in various cartilaginous, ray-finned and lobe-finned fish species. Next, we focused on rainbow trout sAC (rtsAC) and identified 20 potential alternative spliced mRNAs coding for protein isoforms ranging in size from 28 to 186 kDa. Biochemical and kinetic analyses on purified recombinant rtsAC protein determined stimulation by HC O 3   - at physiologically relevant levels for fish internal fluids (EC 50 ∼ 7 mM). rtsAC activity was sensitive to KH7, LRE1, and DIDS (established inhibitors of sAC from other organisms), and insensitive to forskolin and 2,5-dideoxyadenosine (modulators of transmembrane adenylyl cyclases). Western blot and immunocytochemistry revealed high rtsAC expression in gill ion-transporting cells, hepatocytes, red blood cells, myocytes and cardiomyocytes. Analyses in the cell line RTgill-W1 suggested that some of the longer rtsAC isoforms may be preferentially localized in the nucleus, the Golgi apparatus and podosomes. These results indicate that sAC is poised to mediate multiple acid-base homeostatic responses in bony fishes, and provide cues about potential novel functions in mammals.
Competing Interests: The authors declare no competing interests.
(© 2021 The Author(s).)
Databáze: MEDLINE
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