Molecular Characterization of a Novel Intracellular ADP-Ribosyl Cyclase 英文参考文献.docVIP

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Molecular Characterization of a Novel Intracellular ADP-Ribosyl Cyclase 英文参考文献.doc

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Molecular Characterization of a Novel Intracellular ADP-Ribosyl Cyclase 英文参考文献

MolecularCharacterizationofaNovelIntracellular ADP-RibosylCyclase DevChuramani1,MichaelJ.Boulware2.,TimothyJ.Geach3.,AndrewC.R.Martin4,GaryW.Moy5,Yi-HsienSu5,VictorD.Vacquier5, JonathanS.Marchant2,LeslieDale3,SandipPatel1* 1DepartmentofPhysiology,UniversityCollegeLondon,London,UnitedKingdom,2DepartmentofPharmacology,UniversityofMinnesotaMedical School, Minneapolis, Minnesota, United States of America, 3Department of Anatomy and Developmental Biology, University College London, London,UnitedKingdom,4DepartmentofBiochemistryandMolecularBiology,UniversityCollegeLondon,London,UnitedKingdom,5Centerfor MarineBiotechnologyandBiomedicine,ScrippsInstitutionofOceanography,UniversityofCaliforniaatSanDiego,LaJolla,California,UnitedStatesof America Background.ADP-ribosylcyclasesareremarkableenzymescapableofcatalyzingmultiplereactionsincludingthesynthesisof thenovelandpotentintracellularcalciummobilizingmessengers,cyclicADP-riboseandNAADP.NotallADP-ribosylcyclases howeverhavebeencharacterizedatthemolecularlevel.Moreover,thosethathavearelocatedpredominatelyattheoutercell surface and thus away from their cytosolic substrates. Methodology/Principal Findings. Here we report the molecular cloningofanovelexpandedfamilyofADP-ribosylcyclasesfromtheseaurchin,anextensivelyusedmodelorganismforthe studyofinositoltrisphosphate-independentcalciummobilization.Weprovideevidencethatoneoftheisoforms(SpARC1)is asolubleproteinthatistargetedexclusivelytotheendoplasmicreticulumlumenwhenheterologouslyexpressed.Catalytic activityoftherecombinantproteinwasreadilydemonstrableincrudecellhomogenates,evenunderconditionswhereluminal continuitywasmaintained.Conclusions/Significance.OurdatarevealanewintracellularlocationforADP-ribosylcyclases andsuggestthatproductionofcalciummobilizingmessengersmaybecompartmentalized. Citation: Churamani D,BoulwareMJ,GeachTJ,MartinACR,MoyGW,et al (2007)Molecular Characterizationof aNovelIntracellular ADP-Ribosyl Cyclase.PLoSONE2(8):e797.doi:10.1371/journal.pone.0000797 I