title: A defined subset of adenylyl cyclases is regulated by bicarbonate ion.
abstract: The molecular basis by which organisms detect and respond to fluctuations in inorganic carbon is not known. The cyaB1 gene of the cyanobacterium Anabaena sp. PCC7120 codes for a multidomain protein with a C-terminal class III adenylyl cyclase catalyst that was specifically stimulated by bicarbonate ion (EC50 9.6 mm). Bicarbonate lowered substrate affinity but increased reaction velocity. A point mutation in the active site (Lys-646) reduced activity by 95% and was refractory to bicarbonate activation. We propose that Lys-646 specifically coordinates bicarbonate in the active site in conjunction with an aspartate to threonine polymorphism (Thr-721) conserved in class III adenylyl cyclases from diverse eukaryotes and prokaryotes. Using recombinant proteins we demonstrated that adenylyl cyclases that contain the active site threonine (cyaB of Stigmatella aurantiaca and Rv1319c of Mycobacterium tuberculosis) are bicarbonate-responsive, whereas adenylyl cyclases with a corresponding aspartate (Rv1264 of Mycobacterium) are bicarbonate-insensitive. Large numbers of class III adenylyl cyclases may therefore be activated by bicarbonate. This represents a novel mechanism by which diverse organisms can detect bicarbonate ion.
authors.affiliation: Department of Biological and Biomedical Sciences, University of Durham, South Road, Durham, DH1 3LE, United Kingdom. m.j.cann@durham.ac.uk
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Cann
authors.forename: Arne
authors.initials: A
authors.lastname: Hammer
authors.forename: Jie
authors.initials: J
authors.lastname: Zhou
authors.forename: Tobias
authors.initials: T
authors.lastname: Kanacher
mesh_terms: D000262:Adenylyl Cyclases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D019943:Amino Acid Substitution; D017033:Anabaena / Q000201:enzymology* / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001639:Bicarbonates / Q000494:pharmacology*; D001665:Binding Sites; D017124:Conserved Sequence; D004789:Enzyme Activation; D007700:Kinetics; D008239:Lysine; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D017354:Point Mutation; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D001639:Bicarbonates; D000262:Adenylyl Cyclases; D008239:Lysine