ADP ribosyl cyclase activity in rat parotid acinar cells.
Cyclic ADP-ribose is an intracellular compound responsible for Ca2+ release in a wide variety of cell types. It may be implicated in releasing Ca2+ from ryanodine-sensitive pools in exocrine acinar cells. A bifunctional enzyme CD38 can synthesize cADP-ribose and we have characterized its properties by applying a technique in which nicotinamide guanine dinucleotide (NGD+) is used as a substrate for the synthesis of fluorescent cyclic GDP-ribose. This reaction mimics the physiologically relevant reaction in which nicotinamide adenine dinucleotide (NAD+) is converted into non-fluorescent cyclic ADP-ribose. Using NGD+ as a substrate, the reaction shows a half maximal rate of synthesis at 2.6 microM and is competitively inhibited by NAD+ with a k(i) of 12.6 microM. This reveals that both NGD+ and NAD+ are converted by CD38 to their cyclic nucleotides. We have used this fluorescence technique to characterize the extent to which parotid acinar cells contain enzymes capable of synthesizing this class of cyclic nucleotides. We found that after treatment of acinar cells with a detergent which releases intracellular enzymes, NGD+ is converted into its fluorescent derivative with a half maximal rate of synthesis at 16 microM. This reaction is also competitively inhibited by NAD+ with a k(i) of 10 microM. The data indicate that parotid acinar cells contain an enzyme capable of synthesizing the Ca2+ releasing compound, cyclic ADP-ribose. This finding suggests that cyclic ADP-ribose could play a role in Ca2+ release processes from internal stores--an important event in stimulus-secretion coupling.
Looms
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Dept. Medical Physiology, The Panum Institute, University of Copenhagen, Denmark.
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Dissing
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D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D000246:Adenosine Diphosphate Ribose / Q000031:analogs & derivatives* / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D036563:Cyclic ADP-Ribose; D006152:Cyclic GMP / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins; D009244:NAD+ Nucleosidase / Q000378:metabolism*; D010306:Parotid Gland / Q000201:enzymology*; D051381:Rats; D017208:Rats, Wistar
D015703:Antigens, CD; D000943:Antigens, Differentiation; D008562:Membrane Glycoproteins; D036563:Cyclic ADP-Ribose; D000246:Adenosine Diphosphate Ribose; D036541:ADP-ribosyl Cyclase; C498156:Cd38 protein, rat; D009244:NAD+ Nucleosidase; D051997:ADP-ribosyl Cyclase 1; D006152:Cyclic GMP; D002118:Calcium