title: Inhibition of the intrinsic NAD+ glycohydrolase activity of CD38 by carbocyclic NAD analogues.
abstract: Carba-NAD and pseudocarba-NAD are carbocyclic analogues of NAD+ in which a 2,3-dihydroxycyclopentane methanol replaces the beta-d-ribonucleotide ring of the nicotinamide riboside moiety of NAD+ [Slama and Simmons (1988) Biochemistry 27, 183-193]. These carbocyclic NAD+ analogues, related to each other as diastereomers, have been tested as inhibitors of the intrinsic NAD+ glycohydrolase activity of human CD38, dog spleen NAD+ glycohydrolase, mouse CD38 and Aplysia californica cADP-ribose synthetase. Pseudocarba-NAD, the carbocyclic dinucleotide in which l-2,3-dihydroxycyclopentane methanol replaces the d-ribose of the nicotinamide riboside moiety of NAD+, was found to be the more potent inhibitor. Pseudocarba-NAD was shown to inhibit the intrinsic NAD+ glycohydrolase activity of human CD38 competitively, with Ki=148 microM determined for the recombinant extracellular protein domain and Ki=180 microM determined for the native protein expressed as a cell-surface enzyme on cultured Jurkat cells. Pseudocarba-NAD was shown to be a non-competitive inhibitor of the purified dog spleen NAD+ glycohydrolase, with Kis=47 miroM and Kii=198 microM. Neither pseudocarba-NAD nor carba-NAD inhibited mouse CD38 or Aplysia californica cADP-ribose synthetase significantly at concentrations up to 1 mM. The results underscore significant species differences in the sensitivity of these enzymes to inhibition, and indicate that pseudocarba-NAD will be useful as an inhibitor of the enzymic activity of human but not mouse CD38 in studies using cultured cells.
authors.affiliation: Department of Medicinal and Biological Chemistry, College of Pharmacy, University of Toledo, Toledo, OH 43606, USA.
authors.forename: K A
authors.initials: KA
authors.lastname: Wall
authors.forename: M
authors.initials: M
authors.lastname: Klis
authors.forename: J
authors.initials: J
authors.lastname: Kornet
authors.forename: D
authors.initials: D
authors.lastname: Coyle
authors.forename: J C
authors.initials: JC
authors.lastname: Amé
authors.forename: M K
authors.initials: MK
authors.lastname: Jacobson
authors.forename: J T
authors.initials: JT
authors.lastname: Slama
mesh_terms: D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D000943:Antigens, Differentiation / Q000378:metabolism*; D001048:Aplysia / Q000201:enzymology; D002462:Cell Membrane / Q000378:metabolism; D004285:Dogs; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology; D019169:Jurkat Cells; D007700:Kinetics; D008562:Membrane Glycoproteins; D051379:Mice; D009243:NAD / Q000031:analogs & derivatives* / Q000494:pharmacology*; D009244:NAD+ Nucleosidase / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013154:Spleen / Q000201:enzymology; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
chemical_list: D015703:Antigens, CD; D000943:Antigens, Differentiation; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D009243:NAD; C055311:carbanicotinamide adenine dinucleotide; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; C498155:Cd38 protein, mouse; D009244:NAD+ Nucleosidase; D051997:ADP-ribosyl Cyclase 1; D007538:Isoniazid