Hydrolysis of the non-canonical cyclic nucleotide cUMP by PDE9A: kinetics and binding mode.

Autor: Scharrenbroich J; Hannover Medical School, Institute of Pharmacology, Carl-Neuberg-Str. 1, 30625, Hannover, Germany., Kaever V; Research Core Unit Metabolomics, Hannover Medical School, Institute of Pharmacology, Carl-Neuberg-Str. 1, 30625, Hannover, Germany., Dove S; Department of Medicinal Chemistry II, Institute of Pharmacy, University of Regensburg, 93040, Regensburg, Germany., Seifert R; Hannover Medical School, Institute of Pharmacology, Carl-Neuberg-Str. 1, 30625, Hannover, Germany., Schneider EH; Hannover Medical School, Institute of Pharmacology, Carl-Neuberg-Str. 1, 30625, Hannover, Germany. schneider.erich@mh-hannover.de.
Jazyk: angličtina
Zdroj: Naunyn-Schmiedeberg's archives of pharmacology [Naunyn Schmiedebergs Arch Pharmacol] 2019 Feb; Vol. 392 (2), pp. 199-208. Date of Electronic Publication: 2018 Nov 15.
DOI: 10.1007/s00210-018-1582-5
Abstrakt: The non-canonical cyclic nucleotide cUMP and the phosphodiesterase PDE9A both occur in neuronal cells. Using HPLC-coupled tandem mass spectrometry, we characterized the kinetics of PDE9A-mediated cUMP hydrolysis. PDE9A is a low-affinity and high-velocity enzyme for cUMP (V max  = ~ 6 μmol/min/mg; K m  = ~ 401 μM). The PDE9 inhibitor BAY 73-6691 inhibited PDE9A-catalyzed cUMP hydrolysis (K i  = 590 nM). Docking studies indicate two H-bonds between the cUMP uridine moiety and Gln453/Asn405 of PDE9A. By contrast, the guanosine moiety of cGMP forms three H-bonds with Gln453. cCMP is not hydrolyzed at a concentration of 3 μM, but inhibits the PDE9A-catalyzed cUMP hydrolysis at concentrations of 100 μM or more. The probable main reason is that the cytosine moiety cannot act as H-bond acceptor for Gln453. A comparison of PDE9A with PDE7A suggests that the preference of the former for cGMP and cUMP and of the latter for cAMP and cCMP is due to stabilized alternative conformations of the side chain amide of Gln453 and Gln413, respectively. This so-called glutamine switch is known to be involved in the regulation of cAMP/cGMP selectivity of some PDEs.
Databáze: MEDLINE
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