Adenosine Triphosphate Stimulates Aquifex aeolicus MutL Endonuclease Activity 英文参考文献.docVIP

Adenosine Triphosphate Stimulates Aquifex aeolicus MutL Endonuclease Activity 英文参考文献.doc

  1. 1、本文档共9页,可阅读全部内容。
  2. 2、原创力文档(book118)网站文档一经付费(服务费),不意味着购买了该文档的版权,仅供个人/单位学习、研究之用,不得用于商业用途,未经授权,严禁复制、发行、汇编、翻译或者网络传播等,侵权必究。
  3. 3、本站所有内容均由合作方或网友上传,本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺!文档内容仅供研究参考,付费前请自行鉴别。如您付费,意味着您自己接受本站规则且自行承担风险,本站不退款、不进行额外附加服务;查看《如何避免下载的几个坑》。如果您已付费下载过本站文档,您可以点击 这里二次下载
  4. 4、如文档侵犯商业秘密、侵犯著作权、侵犯人身权等,请点击“版权申诉”(推荐),也可以打举报电话:400-050-0827(电话支持时间:9:00-18:30)。
  5. 5、该文档为VIP文档,如果想要下载,成为VIP会员后,下载免费。
  6. 6、成为VIP后,下载本文档将扣除1次下载权益。下载后,不支持退款、换文档。如有疑问请联系我们
  7. 7、成为VIP后,您将拥有八大权益,权益包括:VIP文档下载权益、阅读免打扰、文档格式转换、高级专利检索、专属身份标志、高级客服、多端互通、版权登记。
  8. 8、VIP文档为合作方或网友上传,每下载1次, 网站将根据用户上传文档的质量评分、类型等,对文档贡献者给予高额补贴、流量扶持。如果你也想贡献VIP文档。上传文档
查看更多
Adenosine Triphosphate Stimulates Aquifex aeolicus MutL Endonuclease Activity 英文参考文献

AdenosineTriphosphateStimulatesAquifexaeolicus MutLEndonucleaseActivity JeromeMauris,ThomasC.Evans,Jr* NewEnglandBiolabs,Inc.,Ipswich,Massachusetts,UnitedStatesofAmerica Abstract Background: Human PMS2 (hPMS2) homologues act to nick 59 and 39 to misincorporated nucleotides during mismatch repairinorganismsthatlackMutH.Mn waspreviouslyfoundtostimulatetheendonucleaseactivityofthesehomologues. ++ ATPwasrequiredforthenickingactivityofhPMS2andyPMS1,butwasreportedtoinhibitbacterialMutLproteinsfrom Thermus thermophilus and Aquifex aeolicus that displayed homology to hPMS2. Mutational analysis has identified the DQHA(X)2E(X)4EmotifpresentintheC-terminusofPMS2homologuesasimportantforendonucleaseactivity. Methodologies/PrincipalFindings:WeexaminedtheeffectATPhadontheMn++inducednickingofsupercoiledpBR322 byfull-lengthandmutantA.aeolicusMutL(AaeMutL)proteins.Assaysweresingletimepoint,enzymetitrationexperiments or reaction time courses. The maximum velocity for MutL nicking was determined to be 1.660.0861025s21 and intheabsenceandpresenceofATP,respectively.AMPPNPstimulatedthenickingactivitytoasimilar 4.260.3610 25 21 s extentasATP.AtruncatedAaeMutLproteincomposedofonlytheC-terminal123aminoacidresidueswasfoundtonick supercoiledDNA.Furthermore,mutationsintheconservedC-terminalDQHA(X)2E(X)4EandCPHGRPmotifswereshownto abolishAaeMutLendonucleaseactivity. Conclusions:ATPstimulatedtheMn++inducedendonucleaseactivityofAaeMutL.ExperimentsutilizingAMPPNPimplied thatthestimulationdidnotrequireATPhydrolysis.AmutationintheDQHA(X)2E(X)4EmotifofAaeMutLfurthersupported the role of this region in endonclease activity. For the first time, to our knowledge, we demonstrate that changing the histidineresidueintheconservedCPHGRPmotifabolishesendonucleolyticactivityofahPMS2homologue.Finally,theC- ++ terminal123aminoacidresiduesofAaeMutLweresufficienttodisplayMn inducednickingactivity. Citation: Mauris J, Evans TC Jr (2009) Adenosine Triphosphate Stimulates Aquifex aeolicus MutL Endonucleas

您可能关注的文档

文档评论(0)

1234554321 + 关注
实名认证
文档贡献者

该用户很懒,什么也没介绍

1亿VIP精品文档

相关文档