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Papers In Press, published online ahead of print February 27, 2003
Pharmacology, Weill Medical College of Cornell University, New York, NY 10021
Corresponding Author: jobuck{at}med.cornell.edu
Soluble adenylyl cyclase (sAC) is a widely expressed source of cAMP in mammalian cells which is evolutionarily, structurally, and biochemically distinct from the G protein responsive, transmembrane adenylyl cyclases (tmACs). In contrast to tmACs, sAC is insensitive to heterotrimeric G protein regulation and forskolin stimulation and is uniquely modulated by bicarbonate ions. Here we present the first report detailing kinetic analysis and biochemical properties of purified recombinant sAC. We confirm that bicarbonate regulation is conserved among mammalian sAC orthologs and demonstrate that bicarbonate stimulation is consistent with an increase in the enzymes Vmax with little effect on the apparent Km for substrate, ATP-Mg2+. Bicarbonate can further increase sAC activity by relieving substrate inhibition. We also identify calcium as a direct modulator of sAC activity. In contrast to bicarbonate, calcium stimulates sAC activity by decreasing its apparent Km for ATP-Mg2+. Due to their different mechanisms, calcium and bicarbonate synergistically activate sAC; therefore, small changes of either calcium or bicarbonate will lead to significant changes in cellular cAMP levels.
J. Biol. Chem, 10.1074/jbc.M212475200
Submitted on December 9, 2002
Revised on February 13, 2003
Accepted on February 27, 2003
Kinetic properties of 'soluble' adenylyl cyclase: Synergism between calcium and bicarbonate
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