TY - JOUR
T1 - Metabolism of cyclic ADP-ribose in opossum kidney renal epithelial cells
AU - Beers, K. W.
AU - Chini, E. N.
AU - Hon Cheung Lee, Cheung Lee
AU - Dousa, T. P.
N1 - Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 1995
Y1 - 1995
N2 - We have previously shown that NAD+ inhibits renal Na+-P(i) symport; however, the biochemical mechanism of NAD+ in this action is not clarified. We now propose that NAD+ acts indirectly by first being converted to cyclic ADP-ribose (cADPR), a potent stimulator of intracellular Ca2+ mobilization. In permeabilized opossum kidney (OK) cells, a cell line often employed as a model for study of proximal tubular epithelial transport, cADPR is synthesized from β-NAD+ in a substrate concentration (0.01-1 mM) and time- dependent manner. That cADPR was generated from β-NAD+ by OK cells was verified by coelution with authentic cADPR on anion exchange high-performance liquid chromatography and by homologous desensitization of the Ca2+ release bioassay to authentic cADPR. cADPR synthesized by permeabilized OK cells was not influenced by the addition of parathyroid hormone. The OK cell also contains the enzyme activity necessary to catalyze catabolism of cADPR. Identification of these two key enzyme activities of cADPR metabolism in OK cells is consistent with a possible role of cADPR in regulation of the Na+- P(i) symporter by NAD+ in response to metabolic stimuli.
AB - We have previously shown that NAD+ inhibits renal Na+-P(i) symport; however, the biochemical mechanism of NAD+ in this action is not clarified. We now propose that NAD+ acts indirectly by first being converted to cyclic ADP-ribose (cADPR), a potent stimulator of intracellular Ca2+ mobilization. In permeabilized opossum kidney (OK) cells, a cell line often employed as a model for study of proximal tubular epithelial transport, cADPR is synthesized from β-NAD+ in a substrate concentration (0.01-1 mM) and time- dependent manner. That cADPR was generated from β-NAD+ by OK cells was verified by coelution with authentic cADPR on anion exchange high-performance liquid chromatography and by homologous desensitization of the Ca2+ release bioassay to authentic cADPR. cADPR synthesized by permeabilized OK cells was not influenced by the addition of parathyroid hormone. The OK cell also contains the enzyme activity necessary to catalyze catabolism of cADPR. Identification of these two key enzyme activities of cADPR metabolism in OK cells is consistent with a possible role of cADPR in regulation of the Na+- P(i) symporter by NAD+ in response to metabolic stimuli.
KW - LLC-PK cells
KW - adenosine 5'-diphosphate-ribose
KW - cyclic adenosine 5'-diphosphate-ribose
KW - intracellular calcium
KW - renal epithelia
KW - sodium-inorganic phosphate symport
KW - β-nicotinamide adenine dinucleotide
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U2 - 10.1152/ajpcell.1995.268.3.c741
DO - 10.1152/ajpcell.1995.268.3.c741
M3 - Article
C2 - 7900778
AN - SCOPUS:0028941491
SN - 0363-6143
VL - 268
SP - C741-C746
JO - American Journal of Physiology - Cell Physiology
JF - American Journal of Physiology - Cell Physiology
IS - 3 37-3
ER -