Partially-deuterated samples of HET-s(218–289) fibrils: assignment and deuterium isotope effect

被引:0
作者
Albert A. Smith
Francesco Ravotti
Emilie Testori
Riccardo Cadalbert
Matthias Ernst
Anja Böckmann
Beat H. Meier
机构
[1] ETH Zürich,
[2] Physical Chemistry,undefined
[3] Institut de Biologie et Chimie des Protéines,undefined
[4] Bases Moléculaires et Structurales des Systèmes Infectieux,undefined
[5] Labex Ecofect,undefined
[6] UMR 5086 CNRS,undefined
[7] Université de Lyon,undefined
来源
Journal of Biomolecular NMR | 2017年 / 67卷
关键词
Solid-state NMR; Fibrils; Chemical shift assignment; Deuterium isotope effect; Deuterated proteins; Proton detection;
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学科分类号
摘要
Fast magic-angle spinning and partial sample deuteration allows direct detection of 1H in solid-state NMR, yielding significant gains in mass sensitivity. In order to further analyze the spectra, 1H detection requires assignment of the 1H resonances. In this work, resonance assignments of backbone HN and Hα are presented for HET-s(218–289) fibrils, based on the existing assignment of Cα, Cβ, C’, and N resonances. The samples used are partially deuterated for higher spectral resolution, and the shifts in resonance frequencies of Cα and Cβ due to the deuterium isotope effect are investigated. It is shown that the deuterium isotope effect can be estimated and used for assigning resonances of deuterated samples in solid-state NMR, based on known resonances of the protonated protein.
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页码:109 / 119
页数:10
相关论文
共 204 条
[1]  
Agarwal V(2014)De novo 3D structure determination from sub-milligram protein samples by solid-state 100kHz MAS NMR spectroscopy Angew Chem Int Ed 53 12253-12256
[2]  
Penzel S(2009)Deuterium isotope effects on N-15 backbone chemical shifts in proteins J Biomol NMR 44 119-126
[3]  
Szekely K(2010)High resolution 1H-detected Solid-state NMR spectroscopy of protein aliphatic resonances: access to tertiary structure information J Am Chem Soc 132 15133-15135
[4]  
Cadalbert R(2012)Optimal degree of protonation for 1H detection of aliphatic sites in randomly deuterated proteins as a function of the MAS frequency J Biomol NMR 54 155-168
[5]  
Testori E(2003)Domain organization and structure–function relationship of the HET-s prion protein of Podospora anserina EMBO J 22 2071-2081
[6]  
Oss A(2010)A yeast toxic mutant of HET-s(218–289) prion displays alternative intermediates of amyloidogenesis Biophys J 99 1239-1246
[7]  
Past J(2009)Characterization of different water pools in solid-state NMR protein samples J Biomol NMR 45 319-327
[8]  
Samoson A(2007)Functional amyloid —from bacteria to humans Trends Biochem Sci 32 217-224
[9]  
Ernst M(1998)The use of 2 H, 13 C, 15 N multidimensional NMR to study the structure and dynamics of proteins Annu Rev Biophys Biomol Struct 27 357-406
[10]  
Böckmann A(1997)Global folds of highly deuterated, methyl-protonated proteins by multidimensional NMR Biochemistry 36 1389-1401