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I bastoncelli retinici come cellule altamente specializzate nella fotorecezione : individuazione e caratterizzazione del metabolismo del cADPR
Andrea Fabiano
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Naturwissenschaften, Medizin, Informatik, Technik / Medizin
Beschreibung
Cyclic ADP-ribose (cADPR) is a second messenger modulating intracellular calcium levels. We have previouslydescribed a cADPR-dependent calcium signaling pathway in bovine rod outer segments (ROS), where calcium ions playa pivotal role. ROS ADP-ribosyl cyclase (ADPR-cyclase) was localized in the membrane fraction. In the present work,we examined the properties of the disk ADPR-cyclase through the production of cyclic GDP-ribose from the NAD+analogue NGD+. The enzyme displayed an estimated Km for NGD+ of 12.5 6 0.3 lM, a Vmax of 26.50 6 0.70 pmol cyclicGDP-ribose synthesized/min/mg, and optimal pH of 6.5. The effect of divalent cations (Zn2+, Cu2+, and Ca2+) wasalso tested. Micromolar Zn2+ and Cu2+ inhibited the disk ADPR-cyclase activity (half maximal inhibitory concentration,IC50 5 1.1 and 3.6 lM, respectively). By contrast, Ca2+ ions had no effect. Interestingly, the properties of the intracellularmembrane–associated ROS disk ADPR-cyclase are more similar to those of the ADPR-cyclase found in CD38-deficientmouse brain, than to those of CD38 or CD157. The novel intracellular mammalian ADPR-cyclase would elicit Ca2+ releasefrom the disks at various rates in response to change in free Ca2+ concentrations, caused by light versus dark adaptation, infact there was no difference in disk ADPR-cyclase activity in light or dark conditions. Data suggest that disk ADPR-cyclasemay be a potential target of retinal toxicity of Zn2+ and may shed light to the role of Cu2+ and Zn2+ deficiency in retina
