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Article: Mechanism of cyclizing NAD to cyclic ADP-ribose by ADP-ribosyl cyclase and CD38

TitleMechanism of cyclizing NAD to cyclic ADP-ribose by ADP-ribosyl cyclase and CD38
Authors
Issue Date2009
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
Citation
Journal Of Biological Chemistry, 2009, v. 284 n. 40, p. 27629-27636 How to Cite?
AbstractMammalian CD38 and its Aplysia homolog, ADP-ribosyl cyclase (cyclase), are two prominent enzymes that catalyze the synthesis and hydrolysis of cyclic ADP-ribose (cADPR), a Ca2+ messenger molecule responsible for regulating a wide range of cellular functions. Although both use NAD as a substrate, the cyclase produces cADPR, whereas CD38 produces mainly ADP-ribose (ADPR). To elucidate the catalytic differences and the mechanism of cyclizing NAD, the crystal structure of a stable complex of the cyclase with an NAD analog, ribosyl-2′F-2′-deoxynicotinamide adenine dinucleotide (ribo-2′-F-NAD), was determined. The results show that the analog was a substrate of the cyclase and that during the reaction, the nicotinamide group was released and a stable intermediate was formed. The terminal ribosyl unit at one end of the intermediate formed a close linkage with the catalytic residue (Glu-179), whereas the adenine ring at the other end stacked closely with Phe-174, suggesting that the latter residue is likely to be responsible for folding the linear substrate so that the two ends can be cyclized. Mutating Phe-174 indeed reduced cADPR production but enhanced ADPRproduction, converting the cyclase to be more CD38-like. Changing the equivalent residue in CD38, Thr-221 to Phe, correspondingly enhanced cADPR production, and the double mutation, Thr-221 to Phe and Glu-146 to Ala, effectively converted CD38 to a cyclase. This study provides the first detailed evidence of the cyclization process and demonstrates the feasibility of engineering the reactivity of the enzymes by mutation, setting the stage for the development of tools to manipulate cADPR metabolism in vivo. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.
Persistent Identifierhttp://hdl.handle.net/10722/91953
ISSN
2015 Impact Factor: 4.258
2015 SCImago Journal Rankings: 3.151
PubMed Central ID
ISI Accession Number ID
Funding AgencyGrant Number
National Institutes of HealthGM061568
Hong Kong General Research Fund
National Science Foundation of China/Research Grant Council of Hong Kong (NSFC/RGC) Joint Research Scheme
Funding Information:

This work was supported, in whole or in part, by National Institutes of Health Grant GM061568 (to H.C.L./Q.H.). This work was also supported by grants from the Hong Kong General Research Fund and the National Science Foundation of China/Research Grant Council of Hong Kong (NSFC/RGC) Joint Research Scheme (to H. C. L.).

References

 

DC FieldValueLanguage
dc.contributor.authorGraeff, Ren_HK
dc.contributor.authorLiu, Qen_HK
dc.contributor.authorKriksunov, IAen_HK
dc.contributor.authorKotaka, Men_HK
dc.contributor.authorOppenheimer, Nen_HK
dc.contributor.authorHao, Qen_HK
dc.contributor.authorLee, HCen_HK
dc.date.accessioned2010-09-17T10:31:32Z-
dc.date.available2010-09-17T10:31:32Z-
dc.date.issued2009en_HK
dc.identifier.citationJournal Of Biological Chemistry, 2009, v. 284 n. 40, p. 27629-27636en_HK
dc.identifier.issn0021-9258en_HK
dc.identifier.urihttp://hdl.handle.net/10722/91953-
dc.description.abstractMammalian CD38 and its Aplysia homolog, ADP-ribosyl cyclase (cyclase), are two prominent enzymes that catalyze the synthesis and hydrolysis of cyclic ADP-ribose (cADPR), a Ca2+ messenger molecule responsible for regulating a wide range of cellular functions. Although both use NAD as a substrate, the cyclase produces cADPR, whereas CD38 produces mainly ADP-ribose (ADPR). To elucidate the catalytic differences and the mechanism of cyclizing NAD, the crystal structure of a stable complex of the cyclase with an NAD analog, ribosyl-2′F-2′-deoxynicotinamide adenine dinucleotide (ribo-2′-F-NAD), was determined. The results show that the analog was a substrate of the cyclase and that during the reaction, the nicotinamide group was released and a stable intermediate was formed. The terminal ribosyl unit at one end of the intermediate formed a close linkage with the catalytic residue (Glu-179), whereas the adenine ring at the other end stacked closely with Phe-174, suggesting that the latter residue is likely to be responsible for folding the linear substrate so that the two ends can be cyclized. Mutating Phe-174 indeed reduced cADPR production but enhanced ADPRproduction, converting the cyclase to be more CD38-like. Changing the equivalent residue in CD38, Thr-221 to Phe, correspondingly enhanced cADPR production, and the double mutation, Thr-221 to Phe and Glu-146 to Ala, effectively converted CD38 to a cyclase. This study provides the first detailed evidence of the cyclization process and demonstrates the feasibility of engineering the reactivity of the enzymes by mutation, setting the stage for the development of tools to manipulate cADPR metabolism in vivo. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.en_HK
dc.languageengen_HK
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/en_HK
dc.relation.ispartofJournal of Biological Chemistryen_HK
dc.rightsJournal of Biological Chemistry. Copyright © American Society for Biochemistry and Molecular Biology, Inc.-
dc.subject.meshAntigens, CD38 - chemistry - genetics - metabolism-
dc.subject.meshCalcium - metabolism-
dc.subject.meshCyclic ADP-Ribose - metabolism-
dc.subject.meshMutagenesis, Site-Directed-
dc.subject.meshNAD - metabolism-
dc.titleMechanism of cyclizing NAD to cyclic ADP-ribose by ADP-ribosyl cyclase and CD38en_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0021-9258&volume=284&issue=40&spage=27629&epage=27636&date=2009&atitle=Mechanism+of+Cyclizing+NAD+to+Cyclic+ADP-ribose+by+ADP-ribosyl+Cyclase+and+CD38-
dc.identifier.emailGraeff, R: graeffr@hku.hken_HK
dc.identifier.emailKotaka, M: masayo@hku.hken_HK
dc.identifier.emailHao, Q: qhao@hku.hken_HK
dc.identifier.emailLee, HC: leehc@hku.hken_HK
dc.identifier.authorityGraeff, R=rp01464en_HK
dc.identifier.authorityKotaka, M=rp00293en_HK
dc.identifier.authorityHao, Q=rp01332en_HK
dc.identifier.authorityLee, HC=rp00545en_HK
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1074/jbc.M109.030965en_HK
dc.identifier.pmid19640843-
dc.identifier.pmcidPMC2785691-
dc.identifier.scopuseid_2-s2.0-70350453607en_HK
dc.identifier.hkuros167632-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-70350453607&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume284en_HK
dc.identifier.issue40en_HK
dc.identifier.spage27629en_HK
dc.identifier.epage27636en_HK
dc.identifier.eissn1083-351X-
dc.identifier.isiWOS:000270232300064-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridGraeff, R=7003614053en_HK
dc.identifier.scopusauthoridLiu, Q=35215401600en_HK
dc.identifier.scopusauthoridKriksunov, IA=6507909504en_HK
dc.identifier.scopusauthoridKotaka, M=6604073578en_HK
dc.identifier.scopusauthoridOppenheimer, N=7004639543en_HK
dc.identifier.scopusauthoridHao, Q=7102508868en_HK
dc.identifier.scopusauthoridLee, HC=26642959100en_HK

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