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Article: Acidic residues at the active sites of CD38 and ADP-ribosylt cyclase determine nicotinic acid adenine dinucleotide phosphate (NAADP) synthesis and hydrolysis activities

TitleAcidic residues at the active sites of CD38 and ADP-ribosylt cyclase determine nicotinic acid adenine dinucleotide phosphate (NAADP) synthesis and hydrolysis activities
Authors
Issue Date2006
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, 2006, v. 281 n. 39, p. 28951-28957 How to Cite?
AbstractNicotinic acid adenine dinucleotide phosphate (NAADP) is a novel metabolite of NADP that has now been established as a Ca 2+ messenger in many cellular systems. Its synthesis is catalyzed by multifunctional enzymes, CD38 and ADP-ribosyl cyclase (cyclase). The degradation pathway for NAADP is unknown and no enzyme that can specifically hydrolyze it has yet been identified. Here we show that CD38 can, in fact, hydrolyze NAADP to ADP-ribose 2′-phosphate. This activity was low at neutrality but greatly increased at acidic pH. This novel pH dependence suggests that the hydrolysis is determined by acidic residues at the active site. X-ray crystallography of the complex of CD38 with one of its substrates, NMN, showed that the nicotinamide moiety was in close contact with Glu 146 at 3.27 Å and Asp 155 at 2.52Å. Changing Glu 146 to uncharged Gly and Ala, and Asp 155 to Gln and Asn, by site-directed mutagenesis indeed eliminated the strong pH dependence. Changing Asp 155 to Glu, in contrast, preserved the dependence. The specificity of the two acidic residues was further demonstrated by changing the adjacent Asp 147 to Val, which had minimal effect on the pH dependence. Crystallography confirmed that Asp147 was situated and directed away from the bound substrate. Synthesis of NAADP catalyzed by CD38 is known to have strong preference for acidic pH, suggesting that Glu 146 and Asp 155 are also critical determinants. This was shown to be case by mutagensis. Likewise, using similar approaches, Glu 98 of the cyclase, which is equivalent to Glu 146 in CD38, was found to be responsible for controlling the pH dependence of NAADP synthesis by the cyclase. Based on these findings, a catalytic model is proposed. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.
Persistent Identifierhttp://hdl.handle.net/10722/138602
ISSN
2015 Impact Factor: 4.258
2015 SCImago Journal Rankings: 3.151
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorGraeff, Ren_HK
dc.contributor.authorLiu, Qen_HK
dc.contributor.authorKriksunov, IAen_HK
dc.contributor.authorHao, Qen_HK
dc.contributor.authorHon, CLen_HK
dc.date.accessioned2011-09-02T06:49:08Z-
dc.date.available2011-09-02T06:49:08Z-
dc.date.issued2006en_HK
dc.identifier.citationJournal Of Biological Chemistry, 2006, v. 281 n. 39, p. 28951-28957en_HK
dc.identifier.issn0021-9258en_HK
dc.identifier.urihttp://hdl.handle.net/10722/138602-
dc.description.abstractNicotinic acid adenine dinucleotide phosphate (NAADP) is a novel metabolite of NADP that has now been established as a Ca 2+ messenger in many cellular systems. Its synthesis is catalyzed by multifunctional enzymes, CD38 and ADP-ribosyl cyclase (cyclase). The degradation pathway for NAADP is unknown and no enzyme that can specifically hydrolyze it has yet been identified. Here we show that CD38 can, in fact, hydrolyze NAADP to ADP-ribose 2′-phosphate. This activity was low at neutrality but greatly increased at acidic pH. This novel pH dependence suggests that the hydrolysis is determined by acidic residues at the active site. X-ray crystallography of the complex of CD38 with one of its substrates, NMN, showed that the nicotinamide moiety was in close contact with Glu 146 at 3.27 Å and Asp 155 at 2.52Å. Changing Glu 146 to uncharged Gly and Ala, and Asp 155 to Gln and Asn, by site-directed mutagenesis indeed eliminated the strong pH dependence. Changing Asp 155 to Glu, in contrast, preserved the dependence. The specificity of the two acidic residues was further demonstrated by changing the adjacent Asp 147 to Val, which had minimal effect on the pH dependence. Crystallography confirmed that Asp147 was situated and directed away from the bound substrate. Synthesis of NAADP catalyzed by CD38 is known to have strong preference for acidic pH, suggesting that Glu 146 and Asp 155 are also critical determinants. This was shown to be case by mutagensis. Likewise, using similar approaches, Glu 98 of the cyclase, which is equivalent to Glu 146 in CD38, was found to be responsible for controlling the pH dependence of NAADP synthesis by the cyclase. Based on these findings, a catalytic model is proposed. © 2006 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.titleAcidic residues at the active sites of CD38 and ADP-ribosylt cyclase determine nicotinic acid adenine dinucleotide phosphate (NAADP) synthesis and hydrolysis activitiesen_HK
dc.typeArticleen_HK
dc.identifier.emailGraeff, R: graeffr@hku.hken_HK
dc.identifier.emailHao, Q: qhao@hku.hken_HK
dc.identifier.emailHon, CL: leehc@hku.hken_HK
dc.identifier.authorityGraeff, R=rp01464en_HK
dc.identifier.authorityHao, Q=rp01332en_HK
dc.identifier.authorityHon, CL=rp00545en_HK
dc.description.naturelink_to_OA_fulltexten_US
dc.identifier.doi10.1074/jbc.M604370200en_HK
dc.identifier.pmid16861223-
dc.identifier.scopuseid_2-s2.0-33749409972en_HK
dc.identifier.hkuros134497-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33749409972&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume281en_HK
dc.identifier.issue39en_HK
dc.identifier.spage28951en_HK
dc.identifier.epage28957en_HK
dc.identifier.isiWOS:000240680500049-
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.scopusauthoridHao, Q=7102508868en_HK
dc.identifier.scopusauthoridHon, CL=26642959100en_HK

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