Structural Studies of Sugar Transporters.
糖转运蛋白的结构研究。
基本信息
- 批准号:8663524
- 负责人:
- 金额:$ 11.37万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2014
- 资助国家:美国
- 起止时间:2014-01-10 至 2014-10-31
- 项目状态:已结题
- 来源:
- 关键词:Active Biological TransportAffinityAmino AcidsArginineBindingBinding SitesBiochemicalBiological AssayBradyrhizobiumC-terminalCell membraneCell physiologyCellsChargeComplexCrystallizationDataElectron Spin Resonance SpectroscopyEscherichia coliFamilyFundingGlycogen Storage DiseaseHigh temperature of physical objectHomologous GeneHumanHurricaneInorganic Phosphate TransporterInstitutionKineticsMapsMeasuresMembraneMembrane Transport ProteinsMethodsMolecular ConformationMovementMutagenesisMutateMutationNamesOrganophosphatesPhasePhysiological ProcessesPlayProteinsResolutionRoleRosaSideSiteSpin LabelsStructureSubstrate SpecificitySuspension substanceSuspensionsTechniquesTemperatureTestingVesicleWateralpha-glycerophosphoric acidbiophysical techniquesconformerfallsimprovedmeltingmembernovelreconstitutionresearch studysugar
项目摘要
1. Summary of the original project
Secondary active transport of substrates across the cell membrane is crucial to many cellular and
physiological processes. The largest secondary transporter family is the major facilitator
superfamily (MFS). The sn-glycerol-3-phosphate (G3P) transporter (GlpT) of the inner
membrane of Escherichia coli is a member of the MFS. In the previous funding cycle we
determined the GlpT crystal structure, which suggests a mechanism for substrate translocation
across the membrane that involves a rocker-switch-type movement of the protein. During the
current funding cycle, we aimed to understand the transporter’s substrate specificity, to test the
proposed transport mechanism, and to characterize the conformational changes needed for
substrate transport.
2. Major setback due to Sandy
Most of the original aims of the project were already accomplished, except one part of the
original Aim 4: to get a crystal structure of GlpT in an outward-facing conformation. Part of the
delay in finishing the project was due to the damage that was inflicted onto our institution by
Hurricane Sandy. As of last fall, we finally had 4 Å crystals of a GlpT homolog from
Bradyrhizobium japonicum. As this protein was number 27 among the GlpT homologs we
screened for in our search for a stable protein for crystallization, we named it protein G27.
However, during the hurricane, this campus lost its power and cooling water. As a result, the
temperature in our 18 °C crystallization room rose to 48 °C, causing all our crystals to melt. We
are currently trying to repeat the crystallization experiments.
1.原项目总结
底物在细胞膜上的二次主动运输对许多细胞和
生理过程。最大的次级转运体家族是主要的促进者
超级家族(MFS)。甘油-3-磷酸(G3P)转运蛋白(GlpT)
大肠埃希菌膜是MFS的一员。在上一个资金周期中,我们
确定了GlpT的晶体结构,这表明底物移位的机制
穿过膜,这涉及到蛋白质的摇杆开关式运动。在.期间
当前的资金周期,我们的目标是了解转运蛋白的底物特异性,以测试
建议的运输机制,并表征所需的构象变化
底物运输。
2.桑迪带来的重大挫折
该项目的大部分原定目标已经实现,只有一部分除外
最初的目标4:获得外向型GlpT的晶体结构。部分内容
这个项目延迟完成是因为我们的机构受到了严重的破坏。
飓风桑迪。截至去年秋天,我们终于有了来自
日本慢生根瘤菌。由于该蛋白在GlpT同源物中排在第27位,我们
在寻找稳定的蛋白质进行结晶的过程中,我们将其命名为G27蛋白。
然而,在飓风期间,这个校园失去了电力和冷却水。因此,
我们18°C结晶室的温度升至48°C,导致我们所有的晶体都融化了。我们
目前正在尝试重复结晶实验。
项目成果
期刊论文数量(28)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
ATP binding drives substrate capture in an ECF transporter by a release-and-catch mechanism.
- DOI:10.1038/nsmb.3040
- 发表时间:2015-07
- 期刊:
- 影响因子:16.8
- 作者:Karpowich NK;Song JM;Cocco N;Wang DN
- 通讯作者:Wang DN
Functional characterization of a Na+-dependent dicarboxylate transporter from Vibrio cholerae.
- DOI:10.1085/jgp.201311141
- 发表时间:2014-06
- 期刊:
- 影响因子:0
- 作者:Mulligan C;Fitzgerald GA;Wang DN;Mindell JA
- 通讯作者:Mindell JA
Structure and mechanism of a bacterial sodium-dependent dicarboxylate transporter.
- DOI:10.1038/nature11542
- 发表时间:2012-11-22
- 期刊:
- 影响因子:64.8
- 作者:Mancusso, Romina;Gregorio, G. Glenn;Liu, Qun;Wang, Da-Neng
- 通讯作者:Wang, Da-Neng
Structural biology. Symmetric transporters for asymmetric transport.
- DOI:10.1126/science.1161495
- 发表时间:2008-08-08
- 期刊:
- 影响因子:0
- 作者:Karpowich NK;Wang DN
- 通讯作者:Wang DN
Simple screening method for improving membrane protein thermostability.
- DOI:10.1016/j.ymeth.2011.07.008
- 发表时间:2011-12
- 期刊:
- 影响因子:4.8
- 作者:Mancusso, Romina;Karpowich, Nathan K.;Czyzewski, Bryan K.;Wang, Da-Neng
- 通讯作者:Wang, Da-Neng
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{{ truncateString('DANENG WANG', 18)}}的其他基金
Molecular mechanism of citrate transporter NaCT and its mutations that cause pediatric epilepsies
柠檬酸转运蛋白NaCT及其突变导致小儿癫痫的分子机制
- 批准号:
10393545 - 财政年份:2018
- 资助金额:
$ 11.37万 - 项目类别:
Molecular mechanism of citrate transporter NaCT and its mutations that cause pediatric epilepsies
柠檬酸转运蛋白NaCT及其突变导致小儿癫痫的分子机制
- 批准号:
9904781 - 财政年份:2018
- 资助金额:
$ 11.37万 - 项目类别:
Molecular mechanism of citrate transporter NaCT and its mutations that cause pediatric epilepsies - Revision - 1
柠檬酸转运蛋白 NaCT 的分子机制及其导致小儿癫痫的突变 - 修订版 - 1
- 批准号:
10382590 - 财政年份:2018
- 资助金额:
$ 11.37万 - 项目类别:
Structural Basis of Tetracycline Resistance by Efflux Pump TetL.
外排泵 TetL 的四环素耐药性的结构基础。
- 批准号:
8663548 - 财政年份:2014
- 资助金额:
$ 11.37万 - 项目类别:
Structural and mechanistic studies of INDY proteins
INDY 蛋白的结构和机制研究
- 批准号:
8815304 - 财政年份:2013
- 资助金额:
$ 11.37万 - 项目类别:
Structural and mechanistic studies of INDY proteins
INDY 蛋白的结构和机制研究
- 批准号:
8531420 - 财政年份:2013
- 资助金额:
$ 11.37万 - 项目类别:
Structural and mechanistic studies of INDY proteins
INDY 蛋白的结构和机制研究
- 批准号:
8628114 - 财政年份:2013
- 资助金额:
$ 11.37万 - 项目类别:
Structural basis of tetracycline resistance by efflux pump TetL
外排泵TetL对四环素耐药的结构基础
- 批准号:
7887106 - 财政年份:2010
- 资助金额:
$ 11.37万 - 项目类别:
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