Proximity of Transmembrane Segments 5 and 8 of the Glutamate Transporter GLT-1 Inferred from Paired Cysteine Mutagenesis 英文参考文献.docVIP

下载本文档

13
0
约3.25万字
约 7页
2017-05-13 发布于上海
举报
版权申诉

Proximity of Transmembrane Segments 5 and 8 of the Glutamate Transporter GLT-1 Inferred from Paired Cysteine Mutagenesis 英文参考文献.doc

1、本文档共7页，可阅读全部内容。
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、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、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、该文档为VIP文档，如果想要下载，成为VIP会员后，下载免费。
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、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、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

Proximity of Transmembrane Segments 5 and 8 of the Glutamate Transporter GLT-1 Inferred from Paired Cysteine Mutagenesis 英文参考文献

ProximityofTransmembraneSegments5and8ofthe GlutamateTransporterGLT-1InferredfromPaired CysteineMutagenesis XiupingZhang1,ShaogangQu2* 1China-America Cancer Research Institute, Guangdong Medical College, Dongguan, Guangdong, China, 2Department of Immunology, Southern Medical University, Guangzhou,Guangdong,China Abstract Background: GLT-1 is a glial glutamate transporter which maintains low synaptic concentrations of the excitatory neurotransmitterenablingefficientsynaptictransmission.BasedonthecrystalstructureofthebacterialhomologueGltPh ,it hasbeenproposedthatthereentrantloopHP2,whichconnectstransmembranedomains(TM)7and8,movestoopenand closeaccesstothebindingpocketfromtheextracellularmedium.HowevertheconformationchangebetweenTM5and TM8duringthetransportcycleisnotclearyet.Weusedpairedcysteinemutagenesisinconjunctionwithtreatmentswith Copper(II)(1,10-Phenanthroline)3(CuPh),toverifythepredictedproximityofresidueslocatedatthesestructuralelementsof GLT-1. Methodology/Principal Findings: To assess the proximity of transmembrane domain (TM) 5 relative to TM8 during transportbytheglialglutamatetransporterGLT-1/EAAT2,cysteinepairswereintroducedattheextracellularendsofthese structural elements. A complete inhibition of transport by Copper(II)(1,10-Phenanthroline)3 is observed in the double mutants I295C/I463C and G297C/I463C, but not in the corresponding single mutants. Glutamate and potassium, both expectedtoincreasetheproportionofinward-facingtransporters,significantlyprotectedagainsttheinhibitionoftransport activityofI295C/I463CandG297C/I463CbyCuPh.TransportbythedoublemutantsI295C/I463CandG297C/I463Calsowas 2+ inhibitedbyCd . Conclusions/Significance: Our results suggest that TM5 (Ile-295, Gly-297) is in close proximity to TM8 (Ile-463) in the mammaliantransporter,andthatthespatialrelationshipbetweenthesedomainsisalteredduringthetransportcycle. Citation: Zhang X, Qu S (2011) Proximity of Transmembrane Segments 5 and 8 of the Glutamate Transporter GLT-1 Inferred from Pa