资源描述：
《2005-analytical biochemistry-of cytochrome p450 enzyme activity(参考前言)》由会员上传分享，免费在线阅读，更多相关内容在行业资料-天天文库。
1、ANALYTICALBIOCHEMISTRYAnalyticalBiochemistry344(2005)183–192www.elsevier.com/locate/yabioEvaluationof-cyanoethersasXuorescentsubstratesforassayofcytochromeP450enzymeactivitya,baaaKyung-DonKang,PaulD.Jones,HuazhangHuang,RongZhang,caaLyudmilaA.Mostovich,CraigE.Wheelock,TakahoWatanabe,ca,b,¤LyudmilaF
2、.Gulyaeva,BruceD.HammockaDepartmentofEntomologyandCancerResearchCenter,UniversityofCalifornia,Davis,CA95616,USAbMicrobiologyProgram,UniversityofCalifornia,Davis,CA95616,USAcInstituteofMolecularBiologyandBiophysics,SiberianDivisionofAcademyofMedicalScience,Timakovstr.2,Novosibirsk630117,RussiaReceiv
3、ed7January2005Availableonline11July2005AbstractWehavepreviouslyreportedthesynthesisoffour-cyano-containingethersbasedon2-naphthaldehyde(2-NA)ascytochromeP450(P450)Xuorescentsubstrates.ActivitydetectionwasbasedontheformationofXuorescent2-NAfollowingsubstratehydroly-sis.Amajorlimitationofthesesubstr
4、ateswastheneedtoremoveNADPH,arequiredcofactorforP450oxidation,beforemeasuring2-NAXuorescence.Inthisarticle,wereportthesynthesisofanewseriesofnovelP450substratesusing6-dimethylamino-2-naphtha-ldehyde(6-DMANA),whichhasagreenXuorescentemissionthatiswellseparatedfromtheNADPHspectrum.Amajoradvantageofth
5、e6-DMANAsubstratesisthatNADPHremovalisnotrequiredbeforeXuorescencedetection.Weusedeight-cyanoether-basedsubstratestodeterminetheO-dealkylationactivityofhuman,mouse,andratlivermicrosomes.Inaddition,substrateactivitieswerecomparedwiththecommercialsubstrate7-ethoxyresoruWn(7-ER).Thecatalyticturnoverr
6、atesofboththe6-DMANA-and2-NA-basedsubstrateswereinsomecasesthreefoldfasterthanthecatalyticturnoverrateof7-ER.The2-NA-basedsubstrateshadgreaterturnoverthandidthe6-DMANA-basedsubstrates.MurineandratlivermicrosomespreparedfromanimalsthathadbeentreatedwithvariousP450inducerswereusedtoexamineforisozyme-
7、selectiveturnoverofthesubstrates.Thevastlyimprovedopti-calpropertiesandsyntheticXexibilityofthe-cyanoethercompoundssuggestthattheyarepossiblygoodgeneralP450substrates.Ó2005ElsevierInc.Allrightsreserved.Key
