CAGED CYCLIC ADP-RIBOSE - SYNTHESIS AND USE

Citation
R. Aarhus et al., CAGED CYCLIC ADP-RIBOSE - SYNTHESIS AND USE, The Journal of biological chemistry, 270(13), 1995, pp. 7745-7749
Citations number
22
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology
Journal title
ISSN journal
0021-9258
Volume
270
Issue
13
Year of publication
1995
Pages
7745 - 7749
Database
ISI
SICI code
0021-9258(1995)270:13<7745:CCA-SA>2.0.ZU;2-1
Abstract
Cyclic ADP-ribose (cADPR) is a recently discovered cyclic nucleotide w ith Ca2+ mobilizing activity. Caged cADPR was synthesized by reacting cADPR with 2-nitrophenethyldiazoethane. Elemental analyses, H-1 NMR, a nd extinction coefficient measurements indicate that the product conta ins only one caging group. Anion exchange high pressure liquid chromat ography separated caged cADPR into two forms, which most likely repres ent isomers. Both forms could be uncaged with equal efficiency by UV e xposure to regenerate cADPR. Photolysis of caged cADPR was accomplishe d effectively with a spectrofluorimeter. The efficiency of uncaging de pended on wavelength with UV light shorter than about 320 nm being the most effective. Caged cADPR was biologically inactive and could induc e Ca2+ release from sea urchin egg homogenates only after photolysis. Specificity of the Ca2+ release was shown by inhibition by 8-amino-cAD PR, a specific antagonist of cADPR. To demonstrate its utility in live cells, caged cADPR was microinjected into sea urchin eggs. Photolysis using a mercury light source effectively regenerated cADPR and result ed in Ca2+ mobilization and activation of cortical exocytosis in the e ggs.