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Article: Widespread occurrence in animal tissues of an enzyme catalyzing the conversion of NAD+ into a cyclic metabolite with intracellular Ca2+-mobilizing activity

TitleWidespread occurrence in animal tissues of an enzyme catalyzing the conversion of NAD+ into a cyclic metabolite with intracellular Ca2+-mobilizing activity
Authors
Issue Date1989
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, 1989, v. 264 n. 20, p. 11725-11731 How to Cite?
AbstractA metabolite with intracellular Ca2+-mobilizing activity can be produced by incubating NAD+ with extracts from sea urchin eggs. Structural determination indicates it is a cyclized ADP-ribose, and we have proposed cyclic ADP-ribose as a common name for it. In this study, we addressed the question of how wide-spread is the occurrence of the synthesizing enzyme for this NAD+ metabolite. Incubation of NAD+ with extracts prepared from rabbit liver resulted in a progressive increase in Ca2+ release activity which was monitored by a biological assay using sea urchin egg homogenates. The half-maximal concentration of NAD+ required was about 1 mM. The reaction was stereospecific, and the extracts were sensitive to protease treatment and heat, as well as alkaline pH of about 9.0, indicating the reaction was catalyzed by a protein. The active metabolite was purified by an identical high pressure liquid chromatography (HPLC) procedure used for cyclic ADP-ribose. Functionally, the liver metabolite behaved similarly to cyclic ADP-ribose. Both discharged the same Ca2+ stores in sea urchin egg homogenates with the same half-maximal effective concentrations. Both were active in inducing the cortical exocytosis reaction when microinjected into sea urchin eggs. That they are indeed identical compounds was demonstrated by structural analyses showing that they coeluted on a Partisil 5 SAX HPLC column and had identical 1H NMR spectra. Mass spectrometry indicated a mass of 540.0529 for the molecular ion (M - H)- of the liver metabolite, which was identical to within 0.74 ppm of cyclic ADP-ribose. Furthermore, their collisional activated decomposition mass spectra were virtually superimposable. Extracts from rabbit brain, heart, spleen, and kidney were all active in producing similar Ca2+-releasing metabolites which could be isolated by the same HPLC procedure and had similar elution times on both the mixed mode and the Partisil 5 SAX column. It is therefore apparent that the synthesizing enzyme for cyclic ADP-ribose is a very common enzyme.
Persistent Identifierhttp://hdl.handle.net/10722/171529
ISSN
2015 Impact Factor: 4.258
2015 SCImago Journal Rankings: 3.151
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorRusinko, Nen_US
dc.contributor.authorLee, HCen_US
dc.date.accessioned2012-10-30T06:15:33Z-
dc.date.available2012-10-30T06:15:33Z-
dc.date.issued1989en_US
dc.identifier.citationJournal Of Biological Chemistry, 1989, v. 264 n. 20, p. 11725-11731en_US
dc.identifier.issn0021-9258en_US
dc.identifier.urihttp://hdl.handle.net/10722/171529-
dc.description.abstractA metabolite with intracellular Ca2+-mobilizing activity can be produced by incubating NAD+ with extracts from sea urchin eggs. Structural determination indicates it is a cyclized ADP-ribose, and we have proposed cyclic ADP-ribose as a common name for it. In this study, we addressed the question of how wide-spread is the occurrence of the synthesizing enzyme for this NAD+ metabolite. Incubation of NAD+ with extracts prepared from rabbit liver resulted in a progressive increase in Ca2+ release activity which was monitored by a biological assay using sea urchin egg homogenates. The half-maximal concentration of NAD+ required was about 1 mM. The reaction was stereospecific, and the extracts were sensitive to protease treatment and heat, as well as alkaline pH of about 9.0, indicating the reaction was catalyzed by a protein. The active metabolite was purified by an identical high pressure liquid chromatography (HPLC) procedure used for cyclic ADP-ribose. Functionally, the liver metabolite behaved similarly to cyclic ADP-ribose. Both discharged the same Ca2+ stores in sea urchin egg homogenates with the same half-maximal effective concentrations. Both were active in inducing the cortical exocytosis reaction when microinjected into sea urchin eggs. That they are indeed identical compounds was demonstrated by structural analyses showing that they coeluted on a Partisil 5 SAX HPLC column and had identical 1H NMR spectra. Mass spectrometry indicated a mass of 540.0529 for the molecular ion (M - H)- of the liver metabolite, which was identical to within 0.74 ppm of cyclic ADP-ribose. Furthermore, their collisional activated decomposition mass spectra were virtually superimposable. Extracts from rabbit brain, heart, spleen, and kidney were all active in producing similar Ca2+-releasing metabolites which could be isolated by the same HPLC procedure and had similar elution times on both the mixed mode and the Partisil 5 SAX column. It is therefore apparent that the synthesizing enzyme for cyclic ADP-ribose is a very common enzyme.en_US
dc.languageengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/en_US
dc.relation.ispartofJournal of Biological Chemistryen_US
dc.subject.meshAdenosine Diphosphate Ribose - Metabolismen_US
dc.subject.meshAnimalsen_US
dc.subject.meshCalcium - Metabolismen_US
dc.subject.meshChromatography, High Pressure Liquiden_US
dc.subject.meshLiver - Metabolismen_US
dc.subject.meshMagnetic Resonance Spectroscopyen_US
dc.subject.meshMass Spectrometryen_US
dc.subject.meshNad - Metabolismen_US
dc.subject.meshOvum - Enzymologyen_US
dc.subject.meshRabbitsen_US
dc.subject.meshSea Urchinsen_US
dc.subject.meshSubstrate Specificityen_US
dc.titleWidespread occurrence in animal tissues of an enzyme catalyzing the conversion of NAD+ into a cyclic metabolite with intracellular Ca2+-mobilizing activityen_US
dc.typeArticleen_US
dc.identifier.emailLee, HC:leehc@hku.hken_US
dc.identifier.authorityLee, HC=rp00545en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid2745413-
dc.identifier.scopuseid_2-s2.0-0024381620en_US
dc.identifier.volume264en_US
dc.identifier.issue20en_US
dc.identifier.spage11725en_US
dc.identifier.epage11731en_US
dc.identifier.isiWOS:A1989AF18600032-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridRusinko, N=6508070852en_US
dc.identifier.scopusauthoridLee, HC=26642959100en_US

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