Solid State NMR Studies of Amyloid Proteins
淀粉样蛋白的固态核磁共振研究
基本信息
- 批准号:10687158
- 负责人:
- 金额:$ 70.85万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2017
- 资助国家:美国
- 起止时间:2017-08-01 至 2027-05-31
- 项目状态:未结题
- 来源:
- 关键词:2,4-DinitrophenolAducanumabAlzheimer&aposs DiseaseAlzheimer&aposs disease brainAmino AcidsAmyloidAmyloid FibrilsAmyloid ProteinsAmyloid beta-ProteinAmyloidosisAntibodiesArctic mutationBindingBiological ModelsBrainChemicalsComputer softwareCoupledCouplingCryoelectron MicroscopyDetectionDevelopmentDialysis procedureEarly Onset Alzheimer DiseaseExhibitsFrequenciesGenetic PolymorphismGlobulinsGrantGrowthHydration statusInvestigationIsotopesLabelMagicMeasurementMeasuresMethodologyMethodsModelingMolecular StructureMorphologyN-terminalNMR SpectroscopyPathologicPeptidesPolymorphPopulationPrPProbabilityProcessProteinsPublishingReproducibilityResearchResolutionSamplingSignal TransductionSpectrum AnalysisStructureSystemTailTechniquesTestingThermodynamicsTorsionVariantamyloid peptideamyloid structurebeta-2 Microglobulincatalystexperimental studyflexibilityin vivomethod developmentmutantnext generationparagonsolid state nuclear magnetic resonancesup35toolyeast prion
项目摘要
Project Summary/Abstract
The proposed research focuses on the application and development of magic angle spinning (MAS) NMR
as a tool for structural investigations of amyloid peptides and proteins. The research covers four major
topics.
A. Structure of Amyloid Peptides and Proteins
(1) Aβ1-42 and Aβ1-40: Using 1H detected and DNP magic angle spinning (MAS) and cryoEM we plan the
following experiments on Aβ: (a) a determination of the structure of Aβ1-40; (b) the structure of the
pathologically important plaque seeded Aβ1-42; (c) the structure of the N-terminal tail and the binding of
antibody Aducanumab to the tail; (d) and the structure of mutants for Aβ1-42.
(2) Beta-2-microglobulin (β2m) and DeltaN6-β2m: We plan to determine the structure of the 93 AA β2m-DeltaN6 variant of the dialysis related amyloidosis (DRA) protein. In addition, DeltaN6 is thought to be the
catalyst that seeds β2m plaques in-vivo. We therefore also intend to study mixtures of β2m and DeltaN6.
(3) Amyloid polymorphism: We intend to determine the structure of the three polymorphs of the
amyloidiogenic peptide GNNQQNY from Sup35. These are present in a consistent 1:1:1 population
and will provide the first study of the manner in which defined structural changes alter 13C and 15N
shifts. 1H, 13C and 17O NMR will be used to characterize the structure around the H2O molecules in
the GNNQQNY lattice.
B. NMR methods None of the above structural studies would be possible absent NMR methods to assign
spectra, measure distances and torsion angles, to enhance signal intensities, etc. We therefore plan
to continue the development of the methods essential for these structural investigations. (a) We plan
to continue these experiments by initially preparing U-17O/13C/15N GNNQQNY to further develop the
spectroscopy and then to apply it to spectroscopy of Aβ1-42 and Aβ1-40. The experiments will employ 1H
detection and DNP in order to optimize the sensitivity. PAR and PAIN have been essential to MAS
NMR structure determinations but they are semiquantitative methods to measure 13C-13C and 13C-15N
distances. Using SPINEVOLUTION software and experiments on model systems, we plan to develop
these approaches into quantitative distance measurement techniques.
项目总结/摘要
本文主要研究魔角旋转核磁共振技术的应用和发展
作为淀粉样肽和蛋白质结构研究的工具。该研究涵盖了四个主要领域。
话题
A.淀粉样肽和蛋白质的结构
(1)Aβ1-42和Aβ1-40:使用1H检测和DNP魔角旋转(MAS)和cryoEM,我们计划
对Aβ进行以下实验:(a)Aβ1-40的结构测定;(B)A β1-40的结构测定。
病理上重要的空斑接种Aβ1-42;(c)N-末端尾部的结构和
抗体Aducanumab的尾部;(d)Aβ1-42突变体的结构。
(2)β-2-微球蛋白(β 2 m)和DeltaN 6-β 2 m:我们计划确定透析相关淀粉样变性(AAP)蛋白的93 AA β 2 m-DeltaN 6变体的结构。此外,DeltaN 6被认为是
体内接种β 2 m斑块的催化剂。因此,我们还打算研究β 2 m和DeltaN 6的混合物。
(3)淀粉样蛋白多态性:我们打算确定淀粉样蛋白的三种多晶型物的结构。
来自Sup 35的淀粉样蛋白生成肽GNNQQNY。这些都存在于一个一致的1:1:1人口
并将首次研究确定的结构变化改变13 C和15 N的方式
运动一样的1H、13 C和17 O NMR将用于表征H2O分子周围的结构,
GNNQQNY晶格。
B。如果没有NMR方法,上述结构研究都不可能进行。
光谱,测量距离和扭转角,以增强信号强度等。因此,我们计划
继续发展这些结构调查所必需的方法。(a)我们计划
继续这些实验,首先制备铀-17 O/13 C/15 N GNNQQNY,
并将其应用于Aβ1-42和Aβ1-40的光谱分析。实验将使用1H
检测和DNP,以优化灵敏度。PAR和Pain对于MAS至关重要
NMR结构测定,但它们是测量13 C-13 C和13 C-15 N的半定量方法
的距离.使用SPINEVOLUTION软件和模型系统的实验,我们计划开发
这些方法转化为定量距离测量技术。
项目成果
期刊论文数量(2)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
(17)O NMR Investigation of Water Structure and Dynamics.
(17)水结构和动力学的O NMR研究。
- DOI:10.1021/acs.jpcb.6b05755
- 发表时间:2016
- 期刊:
- 影响因子:0
- 作者:Keeler,EricG;Michaelis,VladimirK;Griffin,RobertG
- 通讯作者:Griffin,RobertG
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ROBERT Guy GRIFFIN其他文献
ROBERT Guy GRIFFIN的其他文献
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{{ truncateString('ROBERT Guy GRIFFIN', 18)}}的其他基金
Q-band Upgrade to an X-band Pulsed EPR spectrometer
Q 波段升级为 X 波段脉冲 EPR 光谱仪
- 批准号:
10177244 - 财政年份:2021
- 资助金额:
$ 70.85万 - 项目类别:
Cryoprobe repairs and amplifier replacement for the 800 and 750 MHz spectrometers
800 和 750 MHz 光谱仪的冷冻探针维修和放大器更换
- 批准号:
10400474 - 财政年份:2019
- 资助金额:
$ 70.85万 - 项目类别:
U.S.-Canada Winter School on Biomolecular Solid State NMR
美国-加拿大生物分子固态核磁共振冬季学校
- 批准号:
9398504 - 财政年份:2017
- 资助金额:
$ 70.85万 - 项目类别:
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