苹果酸脱氢酶的结构及功能-汪新颖

:2008-10-10;:2008-11-13:(1984-),,,,,E-mail:[e-mail];:,,,,,E-mail:[e-mail]。:(30500300);“”(NCET-06-0558);;(06043089);(2005Z032);(2006KJ061A)doi:10.3969/j.issn.1008-9632,2009.04.0691,　1,1,1,2(1.安徽师范大学生物大分子进化重点实验室;2.安徽师范大学分子生物学及生物技术研究所,芜湖241000)　:苹果酸脱氢酶(MDH)可以催化苹果酸与草酰乙酸间的可逆转换,主要参与TCA循环、光合作用、C4循环等代谢途径。苹果酸脱氢酶可分为NAD-依赖性的MDH(NAD-MDH)和NADP-依赖性的MDH(NADP-MDH)。在所有真核生物和大部分细菌中,MDH通常形成同源二聚体,在少数细菌中为四聚体。不同来源的MDH催化机制和它们的动力学性质十分类似,显示了它们具有高度的结构相似性。MDH的功能多样,包括线粒体中的能量提供和植物的活性氧代谢等。回顾了苹果酸脱氢酶在生理学、医学、农学领域的研究进展,并针对其生化特性、空间结构特点、催化机理等生物学功能的分子生物学进展进行了综述。:苹果酸脱氢酶;NAD;NADP;催化机制;功能:Q554.5:B:1008-9632(2009)04-0069-04(malatedehydrogenase,MDH),,,,。MDHs,。,MDHsNAD-MDHs(NAD-MDHs)NADP-MDHs(NADP-MDHs)。NAD-MDHs,。,NAD-MDHs,NADP-MDHs。MDHs,。1　1.1pHMDHspH,8.0,(Streptomycesaureofaciens)MDH(E.coliMDH,EcMDH)pH8.1。MDHs40～65℃。MDHMDH50℃,(BacillusstearothermophilusMDH,BsMDH)65℃。,MDHs,(Aquaspilliumarcticum)MDH(AaMDH)4℃,(Thermophilicbacterium)MDH90℃1h,、。1.2EcMDHMDH。BsMDH。AaMDH,、、、。MDHs。,MDHs,MDHs。,[1,2]。,MDHs。(Chlorobiumtepidum)(Chlorobiumvibrio-forme)MDHs,MDH。(Chloroflexusaurantiacus)MDH,,,[3]。1.36926420098JOURNALOFBIOLOGYVol.26　No.4Aug,2009MDHs,MDH:K+;Mg2+Ca2+;Zn2+、Pb2+Cu2+,Cu2+,1M,97%[4]。MDH(HaloarculamarismortuiMDH,HmMDH),,,,[5]。22.1MDHs:MDHsMDHs(mitMDH),,EcMDHmitMDHmitMDH58.3%47.9%;MDHs,MDH(ThermusflavusMDH,TfMDH)MDHs(chlMDH)MDHs(cyMDH),。MDHs,MDHs。MDHMDH48%～67%。MDHsMDHs,cyMDH、cyMDH59%52%,cyMDHMDH(gMDH)24%,mitMDHcyMDH20%。MDH。(archaeal)MDH(lactatedehydrogenases,LDH),MDHsLDHs,HmMDH,HmMDHLDHsMDHs。MDHsLDHs(paralogous)(geneduplication),3(clade):,,mitMDHscyMDHs;,,LDHs-MDHs(LDH-likeMDH)。LDHsMDHs102,MDHsArg,LDHsGlu。2.2MDHs,MDHs,cyMDHcyM-DH。MDH,MDHs,MDH。NAD-MDHs32～37kDa,NADP-MDHs42kDa。MDH,,,、、。MDHs,NADN-,,β-(Rossman)。4β-α-。MDHsC-。。cyMDH,NAD-MDHsN-NAD、C-,MDH。NAD-MDHs,AspHis,Asp-150His-177EcMDH;Asp-152His-180cyMDH。His-Asp,,MDHsNADHNAD。His、AspMDHs。MDHs2-His-Asp,,2-,,、。2.3NAD-MDHsNADP-MDHs。Asp-53MDH,NAD-MDHs,NADP-MDH,Gly。,MDHs。TakeoTfMDHEX3EX7。EX3TfMDH41、42、45NADP-MDHGly、SerSer。EX77026420098JOURNALOFBIOLOGYVol.26　No.4Aug,20094147Glu-Ile-Pro-Gln-Ala-Met-LysNADP-MDHGly-Ser-Glu-Arg-Ser-Phe-Gln。NAD+NADP+。NADP-MDH84、85、87、88TfMDHGlu-Ile-Glu-Ala,。LDH(Bacillusstearo-thermophilusLDH,BsLDH)EcMDH25%,,。,102,BsLDHGln,EcMDHArg。WilksBsLDHGln-102Arg-102,BsLDHMDH。EcMDHArg-102Gln-102,,。3MDHsNAD(P)+,。MDHs,。--,。,,,His-Asp(1)。1　Fig1　ReactionmechanismofMDH,schemeofNAD-dependentdehydrogenationof2-hydroxyacidsMDHs3Arg。,2Arg,Arg,。cyMDH,Arg-91Arg-97,His-186C2,Arg-161C1,Arg-97C2,。,,,C2,NADH。EcMDH,Arg-81、Arg-87、Arg-153。Arg-81Arg-87,4-2-,Arg-1531-,。4(malatedehydrogenase,MDH),,。MDH,,。MDH:cyMDH,;mitMDHTCA,;MDH;MDH。,。,,,MDH,,[6,7〗。MDH,,。,MDH,D.salinaD.bardwil,MDH,[8]。,F1,MDH,,,,,。10MDHII、、、、,35,MDHII7126420098JOURNALOFBIOLOGYVol.26　No.4Aug,2009[9]。MDH,,MDH,MDH,、。,。[10]。cyMDH,。(Echi-nococcusgranulosus)mitMDH,,[11]。MDH,D,L-D-。D-,4-,、、。D-,D-。,,,、。,MDHs,。MDHs、、、。MDHs、、,MDHs。MDHMDHs。:[1]JaindlM,PoppM.Cyclitolsprotectglutaminesynthetaseandmalatedehydrogenaseagainstheatinduceddeactivationandthermaldenatur-ation[J].BiochemBiophysResCommun,2006,345:761～765.[2]龚　韧,张大伟.猪心苹果酸脱氢酶酶学性质及稳定性研究[J].河南工业大学学报(自然科学版),2007,28(5):42～45.[3]BjorkA,DalhusB,MantzilasD.Stabilizationofatetramericmalatedehydrogenasebyintroductionofadisulfidebridgeatthedimer-dimerinterface[J].JMolBiol,2003,334:811～821.[4]汪少芸,叶秀云.绿豆中苹果酸脱氢酶的性质及生物进化的研究[J].中国食品学报,2006,6(1):262～266.[5]MadernD,EbelC.Influenceofananion-bindingsiteinthestabiliza-tionofhalophilicmalatedehydrogenasefromHaloarculamarismortui[J].Biochimie,2007,89:981～987.[6]王晓云,毕玉芬.植物苹果酸脱氢酶研究进展[J].生物技术通报,2006,4:44～47.[7]赵　胡,陈丽梅.植物有效利用磷素营养有机酸代谢途径基因工程研究进展[J].中国农学通报,2007,23(1):33～37.[8]贺庆华,郑玉才.盐生杜氏藻D.salina和D.bardwil苹果酸脱氢酶基因的克隆和序列分析[J].安徽农业科学,2008,36(11):4435～4436.[9]刘艳荷,陈盛禄.西方蜜蜂苹果酸脱氢酶Ⅱ等位基因频率与经济性状的相关性[J].上海交通大学学报(农业科学版)2006,24(1):79～83.[10]黄　江,胡旭初.牛带绦虫亚洲亚种苹果酸脱氢酶基因的生物信息学分析[J].中国寄生虫学与寄生虫病志,2008,26(3):225～227.[11]张　静,王　洁.细粒棘球蚴(中国大陆株)线粒体苹果酸脱氢酶重组蛋白的表达、纯化及免疫特性分析[J].中国病原生物学杂志,2006,1(4):249～252.StructureandfunctionofmalatedehydrogenasesWANGXin-ying,WANGBo,HOUSong-tao,ZHUGuo-ping(1.TheKeyLaboratoryofMolecularEvolution;2.theInstituteofMolecularBiologyandBiotechnology,AnhuiNormalUniversity,Wuhu241000,China)Abstract:Malatedehydrogenases(MDHs)maycatalyzethereversibleconversionbetweenmalateandoxaloacetate,whichplayakeyroleinmanymetabolicpathwayssuchasTCAcycle,photosynthesis,C4cycleandsoon.MDHsaredividedintoNAD-dependentMDHsandNADP-dependentMDHs,whichusuallyarehomodimericmoleculesinmostorganisms,includingalleukaryotesandthemostbacterialspecies(somebacterialMDHsaretetramer).Theenzymesshareacommoncatalyticmechanismandtheirkineticprop-ertiesaresimilar,whichdemonstrateahighdegreeofstructuralsimilarity.ThefunctionsofMDHsarebiologicaldiversity,includingenergygenerationinmitochondrial,reactiveoxygenspeciesmetabolisminplants,etc.Basedonthebiochemistry,stereoconformationandcatalysismechanism,theadvancesofMDHsinmolecularbiology,physiology,medicineandagriculturearediscussed.Keywords:malatedehydrogenase;NAD;NADP;catalysismechanism;function7226420098JOURNALOFBIOLOGYVol.26　No.4Aug,2009