Mechanistic Implications of Cyclic ADP-Ribose Hydrolysis and Methanolysis Catalyzed by Calf Spleen NAD+Glycohydrolase

1994 ◽  
Vol 204 (3) ◽  
pp. 1279-1285 ◽  
Author(s):  
H. Mullersteffner ◽  
M. Muzard ◽  
N. Oppenheimer ◽  
F. Schuber
Keyword(s):  
Nad Glycohydrolase ◽  
Cyclic Adp Ribose

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).

scholarly journals Expression of cyclic ADP-ribose-synthetizing CD38 molecule on human platelet membrane

Blood ◽  
1996 ◽  
Vol 87 (6) ◽  
pp. 2308-2313 ◽  
Author(s):  
G Ramaschi ◽  
M Torti ◽  
ET Festetics ◽  
F Sinigaglia ◽  
F Malavasi ◽  
...  
Keyword(s):  
Cellular Localization ◽  
Human Platelet ◽  
Human Platelets ◽  
Enzymatic Activities ◽  
Cell Surface Molecule ◽  
Nad Glycohydrolase ◽  
Cd38 Expression ◽  
Cyclic Adp Ribose ◽  
A Cell ◽  
Hydrolysis Of

CD38 is a cell surface molecule widely used as a marker for immature and activated lymphocytes. It has been recently shown that CD38 displays three enzymatic activities: hydrolysis of NAD+ to ADP-ribose, synthesis of cyclic ADP-ribose from NAD+, and hydrolysis of cyclic ADP- ribose to ADP-ribose. Thus, CD38 plays a key role in the synthesis of cyclic ADP-ribose, a calcium-mobilizing compound. We investigate here the expression and cellular localization of CD38 in human platelets using a specific monoclonal antibody. Results showed that CD38 is expressed by human platelet membranes. Moreover, we show that platelet CD38 possesses NAD glycohydrolase, ADP-ribose cyclase, and cyclic ADP- ribose hydrolase activities. This finding indicates that the calcium- mobilizing agent cyclic ADP-ribose can be synthetized by human platelets and raises the question about the possible role of CD38 expression and enzymatic activities in the signal transduction pathways leading to platelet activation.

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 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 A Single Residue at the Active Site of CD38 Determines Its NAD Cyclizing and Hydrolyzing Activities

2001 ◽  
Vol 276 (15) ◽  
pp. 12169-12173 ◽  
Author(s):  
Richard Graeff ◽  
Cyrus Munshi ◽  
Robert Aarhus ◽  
Malcolm Johns ◽  
Hon Cheung Lee
Keyword(s):  
Active Site ◽  
Structural Changes ◽  
Site Directed Mutagenesis ◽  
Cyclase Activity ◽  
Cyclization Reactions ◽  
Base Exchange ◽  
Nad Glycohydrolase ◽  
Cyclic Adp Ribose ◽  
Conserved Residue ◽  
Hydrolysis Activity

CD38 is a multifunctional enzyme involved in metabolizing two Ca2+messengers, cyclic ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate (NAADP). When incubated with NAD, CD38 predominantly hydrolyzes it to ADP-ribose (NAD glycohydrolase), but a trace amount of cADPR is also produced through cyclization of the substrate. Site-directed mutagenesis was used to investigate the amino acid important for controlling the hydrolysis and cyclization reactions. CD38 and its mutants were produced in yeast, purified, and characterized by immunoblot. Glu-146 is a conserved residue present in the active site of CD38. Its replacement with Phe greatly enhanced the cyclization activity to a level similar to that of the NAD hydrolysis activity. A series of additional replacements was made at the Glu-146 position including Ala, Asn, Gly, Asp, and Leu. All the mutants exhibited enhanced cyclase activity to various degrees, whereas the hydrolysis activity was inhibited greatly. E146A showed the highest cyclase activity, which was more than 3-fold higher than its hydrolysis activity. All mutants also cyclized nicotinamide guanine dinucleotide to produce cyclic GDP. This activity was enhanced likewise, with E146A showing more than 9-fold higher activity than the wild type. In addition to NAD, CD38 also hydrolyzed cADPR effectively, and this activity was correspondingly depressed in the mutants. When all the mutants were considered, the two cyclase activities and the two hydrolase activities were correlated linearly. The Glu-146 replacements, however, only minimally affected the base-exchange activity that is responsible for synthesizing NAADP. Homology modeling was used to assess possible structural changes at the active site of E146A. These results are consistent with Glu-146 being crucial in controlling specifically and selectively the cyclase and hydrolase activities of CD38.

scholarly journals NAD+-glycohydrolase fromStreptococcus pyogenesshows cyclic ADP-ribose forming activity

1995 ◽  
Vol 130 (2-3) ◽  
pp. 201-204 ◽  
Author(s):  
Tadahiro Karasawa ◽  
Shin Takasawa ◽  
Kiyotaka Yamakawa ◽  
Hideto Yonekura ◽  
Hiroshi Okamoto ◽  
...  
Keyword(s):  
Nad Glycohydrolase ◽  
Cyclic Adp Ribose
Sign in / Sign up

Export Citation Format

Share Document