scholarly journals Identification of bovine liver mitochondrial NAD+ glycohydrolase as ADP-ribosyl cyclase

1997 ◽  
Vol 326 (2) ◽  
pp. 401-405 ◽  
Author(s):  
Mathias ZIEGLER ◽  
Dierk JORCKE ◽  
Manfred SCHWEIGER
Keyword(s):  
Calcium Release ◽  
Bovine Liver ◽  
Enzymic Activity ◽  
Cyclase Activity ◽  
Partial Recovery ◽  
Nad Glycohydrolase ◽  
Adp Ribosyl Cyclase ◽  
Cyclic Adp Ribose ◽  
Hydrolysis Of ◽  
Synthesis And Degradation

The present investigation identifies bovine liver mitochondrial NADase (NAD+ glycohydrolase) as a member of the class of bifunctional ADP-ribosyl cyclases/cyclic ADP-ribose hydrolases, known to be potential second messenger enzymes. These enzymes catalyse the synthesis and degradation of cyclic ADP-ribose, a potent intracellular calcium-mobilizing agent. The mitochondrial enzyme utilized the NAD+ analogues nicotinamide guanine dinucleotide (NGD+) and nicotinamide hypoxanthine dinucleotide (NHD+) to form fluorescent cyclic purine nucleoside diphosphoriboses. ADP-ribosyl cyclase activity was also demonstrated using 32P-labelled NAD+ as substrate. The identity of NADase and ADP-ribosyl cyclase was supported by their co-migration in SDS/polyacrylamide gels. Cyclase activity was visualized directly within the gel by detecting the formation of fluorescent cyclic IDP-ribose from NHD+. The enzyme catalysed the hydrolysis of cyclic ADP-ribose to ADP-ribose. Moreover, in the presence of nicotinamide and cyclic ADP-ribose the enzyme synthesized NAD+. Both the ADP-ribosyl cyclase and NADase activities of the enzyme were strongly inhibited by reducing agents. Treatment of the NADase with dithiothreitol caused the apparent inactivation of the enzyme. Subsequent removal of the reducing agent and addition of oxidized glutathione led to a partial recovery of enzymic activity. The results support a model for pro-oxidant-induced calcium release from mitochondria involving cyclic ADP-ribose as a specific messenger, rather than the non-enzymic modification of proteins by ADP-ribose.

scholarly journals Nicotinamide inhibits cyclic ADP-ribose-mediated calcium signalling in sea urchin eggs

1996 ◽  
Vol 319 (2) ◽  
pp. 613-617 ◽  
Author(s):  
Jaswinder K SETHI ◽  
Ruth M EMPSON ◽  
Antony GALIONE
Keyword(s):  
Sea Urchin ◽  
Calcium Release ◽  
Calcium Signalling ◽  
Hydrolytic Activity ◽  
Calcium Release Channels ◽  
Adp Ribosyl Cyclase ◽  
Sea Urchin Egg ◽  
Cyclic Adp Ribose ◽  
Single Polypeptide ◽  
Hydrolysis Of

Cyclic ADP ribose (cADPR) is a potent Ca2+-releasing agent, and putative second messenger, the endogenous levels of which are tightly regulated by synthetic (ADP-ribosyl cyclases) and degradative (cADPR hydrolase) enzymes. These enzymes have been characterized in a number of mammalian and invertebrate tissues and their activities are often found on a single polypeptide. β-NAD+, cGMP and nitric oxide (NO) have been reported to mobilize Ca2+ in the sea urchin egg via the cADPR-mediated pathway. We now report that in sea urchin egg homogenates, nicotinamide inhibits the Ca2+-mobilizing action of β-NAD+, cGMP and NO, but has no effect on cADPR-induced Ca2+ release. Moreover, nicotinamide inhibits cGMP-induced regenerative Ca2+ waves in the intact sea urchin egg. By successfully separating the cADPR-metabolizing machinery from that which releases Ca2+, we have shown that nicotinamide inhibits cADPR-mediated Ca2+ signalling at the level of cADPR generation. Importantly, nicotinamide had no effect upon the hydrolysis of cADPR, and its selective action on cyclase activity was supported by its inhibition of purified Aplysia ADP-ribosyl cyclase, which does not exhibit detectable hydrolytic activity. The action of nicotinamide in blocking Ca2+ release by β-NAD+, cGMP and NO strongly suggests that these agents act as modulators of cADPR synthesis rather than to sensitize calcium release channels to cADPR.

scholarly journals CD38-dependent ADP-ribosyl cyclase activity in developing and adult mouse brain

2003 ◽  
Vol 370 (1) ◽  
pp. 175-183 ◽  
Author(s):  
Claire CENI ◽  
Nathalie POCHON ◽  
Virginie BRUN ◽  
Hélène MULLER-STEFFNER ◽  
Annie ANDRIEUX ◽  
...  
Keyword(s):  
Developmental Stages ◽  
The Body ◽  
Adult Brain ◽  
Cyclase Activity ◽  
Wild Type ◽  
Intracellular Location ◽  
Nad Glycohydrolase ◽  
Adp Ribosyl Cyclase ◽  
Activity Measurements ◽  
Cyclic Adp Ribose

CD38 is a transmembrane glycoprotein that is expressed in many tissues throughout the body. In addition to its major NAD+-glycohydrolase activity, CD38 is also able to synthesize cyclic ADP-ribose, an endogenous calcium-regulating molecule, from NAD+. In the present study, we have compared ADP-ribosyl cyclase and NAD+-glycohydrolase activities in protein extracts of brains from developing and adult wild-type and Cd38-/- mice. In extracts from wild-type brain, cyclase activity was detected spectrofluorimetrically, using nicotinamide—guanine dinucleotide as a substrate (GDP-ribosyl cyclase activity), as early as embryonic day 15. The level of cyclase activity was similar in the neonate brain (postnatal day 1) and then increased greatly in the adult brain. Using [14C]NAD+ as a substrate and HPLC analysis, we found that ADP-ribose is the major product formed in the brain at all developmental stages. Under the same experimental conditions, neither NAD+-glycohydrolase nor GDP-ribosyl cyclase activity could be detected in extracts of brains from developing or adult Cd38-/- mice, demonstrating that CD38 is the predominant constitutive enzyme endowed with these activities in brain at all developmental stages. The activity measurements correlated with the level of CD38 transcripts present in the brains of developing and adult wild-type mice. Using confocal microscopy we showed, in primary cultures of hippocampal cells, that CD38 is expressed by both neurons and glial cells, and is enriched in neuronal perikarya. Intracellular NAD+-glycohydrolase activity was measured in hippocampal cell cultures, and CD38-dependent cyclase activity was higher in brain fractions enriched in intracellular membranes. Taken together, these results lead us to speculate that CD38 might have an intracellular location in neural cells in addition to its plasma membrane location, and may play an important role in intracellular cyclic ADP-ribose-mediated calcium signalling in brain tissue.

scholarly journals Snake venom NAD glycohydrolases: primary structures, genomic location, and gene structure

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6154 ◽  
Author(s):  
Ivan Koludarov ◽  
Steven D. Aird
Keyword(s):  
Snake Venom ◽  
Active Site ◽  
Disulfide Bonds ◽  
Venom Gland ◽  
Catalytic Residue ◽  
Nad Glycohydrolase ◽  
Adp Ribosyl Cyclase ◽  
Genomic Location ◽  
Cyclic Adp Ribose ◽  
Very High

NAD glycohydrolase (EC 3.2.2.5) (NADase) sequences have been identified in 10 elapid and crotalid venom gland transcriptomes, eight of which are complete. These sequences show very high homology, but elapid and crotalid sequences also display consistent differences. As in Aplysia kurodai ADP-ribosyl cyclase and vertebrate CD38 genes, snake venom NADase genes comprise eight exons; however, in the Protobothrops mucrosquamatus genome, the sixth exon is sometimes not transcribed, yielding a shortened NADase mRNA that encodes all six disulfide bonds, but an active site that lacks the catalytic glutamate residue. The function of this shortened protein, if expressed, is unknown. While many vertebrate CD38s are multifunctional, liberating both ADP-ribose and small quantities of cyclic ADP-ribose (cADPR), snake venom CD38 homologs are dedicated NADases. They possess the invariant TLEDTL sequence (residues 144–149) that bounds the active site and the catalytic residue, Glu228. In addition, they possess a disulfide bond (Cys121–Cys202) that specifically prevents ADP-ribosyl cyclase activity in combination with Ile224, in lieu of phenylalanine, which is requisite for ADPR cyclases. In concert with venom phosphodiesterase and 5′-nucleotidase and their ecto-enzyme homologs in prey tissues, snake venom NADases comprise part of an envenomation strategy to liberate purine nucleosides, and particularly adenosine, in the prey, promoting prey immobilization via hypotension and paralysis.

scholarly journals Human CD38 is an authentic NAD(P)+ glycohydrolase

1998 ◽  
Vol 330 (3) ◽  
pp. 1383-1390 ◽  
Author(s):  
Valérie BERTHELIER ◽  
Jean-Michel TIXIER ◽  
Hélène MULLER-STEFFNER ◽  
Francis SCHUBER ◽  
Philippe DETERRE
Keyword(s):  
Molecular Mechanisms ◽  
Reaction Scheme ◽  
Reaction Pathways ◽  
Catalytic Activities ◽  
Nad Glycohydrolase ◽  
Cyclization Process ◽  
Adp Ribosyl Cyclase ◽  
Cyclic Adp Ribose ◽  
Water Hydrolysis ◽  
Low Efficiency

The leucoyte surface antigen CD38 has been shown to be an ecto-enzyme with multiple catalytic activities. It is principally a NAD+ glycohydrolase that transforms NAD+ into ADP-ribose and nicotinamide. CD38 is also able to produce small amounts of cyclic ADP-ribose (ADP-ribosyl cyclase activity) and to hydrolyse this cyclic metabolite into ADP-ribose (cyclic ADP-ribose hydrolase activity). To classify CD38 among the enzymes that transfer the ADP-ribosyl moiety of NAD+ to a variety of acceptors, we have investigated its substrate specificity and some characteristics of its kinetic and molecular mechanisms. We find that CD38-catalysed cleavage of the nicotinamide-ribose bond results in the formation of an E·ADP-ribosyl intermediary complex, which is common to all reaction pathways; this intermediate reacts (1) with acceptors such as water (hydrolysis), methanol (methanolysis) or pyridine (transglycosidation), and (2) intramolecularly, yielding cyclic ADP-ribose with a low efficiency. This reaction scheme is also followed when using nicotinamide guanine dinucleotide as an alternative substrate; in this case, however, the cyclization process is highly favoured. The results obtained here are not compatible with the prevailing model for the mode of action of CD38, according to which this enzyme produces first cyclic ADP-ribose which is then immediately hydrolysed into ADP-ribose (i.e. sequential ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase activities). We show instead that the cyclic metabolite was a reaction product of CD38 rather than an obligatory reaction intermediate during the glycohydrolase activity. Altogether our results lead to the conclusion that CD38 is an authentic ‘classical’ NAD(P)+ glycohydrolase (EC 3.2.2.6).

Regulation of store-operated Ca2+ entry by CD38 in human airway smooth muscle

2008 ◽  
Vol 294 (2) ◽  
pp. L378-L385 ◽  
Author(s):  
Gary C. Sieck ◽  
Thomas A. White ◽  
Michael A. Thompson ◽  
Christina M. Pabelick ◽  
Mark E. Wylam ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Airway Smooth Muscle ◽  
Critical Role ◽  
Human Airway Smooth Muscle ◽  
Cd38 Expression ◽  
Adp Ribosyl Cyclase ◽  
Cyclic Adp Ribose ◽  
Intracellular Ca ◽  
Interfering Rna ◽  
Synthesis And Degradation

The ectoenzyme CD38 catalyzes synthesis and degradation of cyclic ADP ribose in airway smooth muscle (ASM). The proinflammatory cytokine TNFα, which enhances agonist-induced intracellular Ca2+ ([Ca2+]i) responses, has been previously shown to increases CD38 expression. In the present study, we tested the hypothesis that the effects of TNFα on CD38 expression vs. changes in [Ca2+]i regulation in ASM cells are linked. Using isolated human ASM cells, CD38 expression was either increased (transfection) or knocked down [small interfering RNA (siRNA)], and [Ca2+]i responses to sarcoplasmic reticulum depletion [i.e., store-operated Ca2+ entry (SOCE)] were evaluated in the presence vs. absence of TNFα. Results confirmed that TNFα significantly increased CD38 expression and ADP-ribosyl cyclase activity, an effect inhibited by CD38 siRNA, but unaltered by CD38 overexpression. CD38 suppression blunted, whereas overexpression enhanced, ACh-induced [Ca2+]i responses. TNFα-induced enhancement of [Ca2+]i response to agonist was blunted by CD38 suppression, but enhanced by CD38 overexpression. Finally, TNFα-induced increase in SOCE was blunted by CD38 siRNA and potentiated by CD38 overexpression. Overall, these results indicate a critical role for CD38 in TNFα-induced enhancement of [Ca2+]i in human ASM cells, and potentially to TNFα augmentation of airway responsiveness.

scholarly journals Oscillation of ADP-ribosyl cyclase activity during the cell cycle and function of cyclic ADP-ribose in a unicellular organism, Euglena gracilis

FEBS Letters ◽  
1997 ◽  
Vol 405 (1) ◽  
pp. 104-106 ◽  
Author(s):  
Wataru Masuda ◽  
Shigeo Takenaka ◽  
Kiyoshi Inageda ◽  
Hiroshi Nishina ◽  
Katsunobu Takahashi ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Euglena Gracilis ◽  
Cyclase Activity ◽  
Unicellular Organism ◽  
Adp Ribosyl Cyclase ◽  
Cyclic Adp Ribose ◽  
And Function

scholarly journals cADP-ribose formation by blood platelets is not responsible for intracellular calcium mobilization

1998 ◽  
Vol 331 (2) ◽  
pp. 431-436 ◽  
Author(s):  
Philippe OHLMANN ◽  
Claude LERAY ◽  
Catherine RAVANAT ◽  
Adel HALLIA ◽  
Dominique CASSEL ◽  
...  
Keyword(s):  
Intracellular Calcium ◽  
Plasma Membranes ◽  
Calcium Release ◽  
Blood Platelets ◽  
Human Platelets ◽  
Cyclase Activity ◽  
Dependent Manner ◽  
Intact Cells ◽  
Adp Ribosyl Cyclase ◽  
Intracellular Calcium Mobilization

Human platelet CD38 is a multifunctional ectoenzyme catalysing the synthesis and hydrolysis of cADP-ribose (cADPR), a recently identified calcium-mobilizing agent that acts independently of d-myo-inositol 1,4,5-trisphosphate and is known to be expressed by human platelets. The present work shows that ADP-ribosyl cyclase activity is exclusively a membrane activity, of which the major part is located in plasma membranes and a small part in internal membranes. In broken cells, cyclase activity was insensitive to the presence of calcium and was not modulated by agonists such as thrombin or ADP, whereas in intact cells thrombin increased cADPR formation by 30%, an effect due to fusion of granules with the plasma membrane. In order to assess the role of cADPR as a calcium-mobilizing agent, vesicles were prepared from internal membranes and loaded with 45CaCl2. These vesicles were efficiently discharged by IP3 in a dose-dependent manner, but were not responsive to cADPR or ryanodine in the presence or absence of calmodulin. Thus cADPR is unlikely to play a role in intracellular calcium release in human blood platelets.

Catalytic properties of the retinal rod outer segment disk ADP-ribosyl cyclase

2011 ◽  
Vol 28 (2) ◽  
pp. 121-128 ◽  
Author(s):  
ANDREA FABIANO ◽  
ISABELLA PANFOLI ◽  
DANIELA CALZIA ◽  
MAURIZIO BRUSCHI ◽  
SILVIA RAVERA ◽  
...  
Keyword(s):  
Divalent Cations ◽  
Cyclase Activity ◽  
The Novel ◽  
Intracellular Membrane ◽  
Adp Ribosyl Cyclase ◽  
Retinal Rod ◽  
Cyclic Adp Ribose ◽  
Response To Change ◽  
Shed Light

AbstractCyclic ADP-ribose (cADPR) is a second messenger modulating intracellular calcium levels. We have previously described a cADPR-dependent calcium signaling pathway in bovine rod outer segments (ROS), where calcium ions play a 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 ± 0.3 μM, a Vmax of 26.50 ± 0.70 pmol cyclic GDP-ribose synthesized/min/mg, and optimal pH of 6.5. The effect of divalent cations (Zn2+, Cu2+, and Ca2+) was also tested. Micromolar Zn2+ and Cu2+ inhibited the disk ADPR-cyclase activity (half maximal inhibitory concentration, IC50 = 1.1 and 3.6 μM, respectively). By contrast, Ca2+ ions had no effect. Interestingly, the properties of the intracellular membrane–associated ROS disk ADPR-cyclase are more similar to those of the ADPR-cyclase found in CD38-deficient mouse brain, than to those of CD38 or CD157. The novel intracellular mammalian ADPR-cyclase would elicit Ca2+ release from the disks at various rates in response to change in free Ca2+ concentrations, caused by light versus dark adaptation, in fact there was no difference in disk ADPR-cyclase activity in light or dark conditions. Data suggest that disk ADPR-cyclase may be a potential target of retinal toxicity of Zn2+ and may shed light to the role of Cu2+ and Zn2+ deficiency in retina.

scholarly journals Regulation of NAD+ glycohydrolase activity by NAD+-dependent auto-ADP-ribosylation

1996 ◽  
Vol 318 (3) ◽  
pp. 903-908 ◽  
Author(s):  
Myung-Kwan HAN ◽  
Ji-Young LEE ◽  
Yee-Sook CHO ◽  
Young M SONG ◽  
Nyeon-Hyoung AN ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Physiological Role ◽  
Enzymic Activity ◽  
Adp Ribosylation ◽  
Nad Glycohydrolase ◽  
Intact Rabbit ◽  
Hydrolysis Of

NAD+ glycohydrolase (NADase; EC 3.2.2.5) is an enzyme that catalyses hydrolysis of NAD+ to produce ADP-ribose and nicotinamide. Its physiological role and the regulation of its enzymic activity have not been fully elucidated. In the present study, the mechanism of self-inactivation of NADase by its substrate, NAD+, was investigated by using intact rabbit erythrocytes and purified NADase. Our results suggest that inactivation of NADase was due an auto-ADP-ribosylation reaction. ADP-ribosylated NADase of rabbit erythrocytes was deADP-ribosylated when incubated without NAD+, and thus enzyme activity was simultaneously restored. These findings suggest that reversible auto-ADP-ribosylation of NADase might regulate the enzyme's activity in vivo.

Unifying mechanism for Aplysia ADP-ribosyl cyclase and CD38/NAD+ glycohydrolases

2000 ◽  
Vol 349 (1) ◽  
pp. 203-210 ◽  
Author(s):  
Céline CAKIR-KIEFER ◽  
Hélène MULLER-STEFFNER ◽  
Francis SCHUBER
Keyword(s):  
Reaction Pathway ◽  
Reaction Scheme ◽  
Hydrolysis Rate ◽  
Cleavage Rate ◽  
Sequential Reaction ◽  
Nad Glycohydrolase ◽  
Adp Ribosyl Cyclase ◽  
High Concentrations ◽  
Hydrolysis Of ◽  
Competing Pathways

Highly purified Aplysia californica ADP-ribosyl cyclase was found to be a multifunctional enzyme. In addition to the known transformation of NAD+ into cADP-ribose this enzyme is able to catalyse the solvolysis (hydrolysis and methanolysis) of cADP-ribose. This cADP-ribose hydrolase activity, which becomes detectable only at high concentrations of the enzyme, is amplified with analogues such as pyridine adenine dinucleotide, in which the cleavage rate of the pyridinium-ribose bond is much reduced compared with NAD+. Although the specificity ratio Vmax/Km is in favour of NAD+ by 4 orders of magnitude, this multifunctionality allowed us to propose a ‘partitioning’ reaction scheme for the Aplysia enzyme, similar to that established previously for mammalian CD38/NAD+ glycohydrolases. This mechanism involves the formation of a single oxocarbenium-type intermediate that partitions to cADP-ribose and solvolytic products via competing pathways. In favour of this mechanism was the finding that the enzyme also catalysed the hydrolysis of NMN+, a substrate that cannot undergo cyclization. The major difference between the mammalian and the invertebrate enzymes resides in their relative cyclization/hydrolysis rate-constant ratios, which dictate their respective yields of cADP-ribose (ADP-ribosyl cyclase activity) and ADP-ribose (NAD+ glycohydrolase activity). For the Aplysia enzyme's catalysed transformation of NAD+ we favour a mechanism where the formation of cADP-ribose precedes that of ADP-ribose; i.e. macroscopically the invertebrate ADP-ribosyl cyclase conforms to a sequential reaction pathway as a limiting form of the partitioning mechanism.

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