<sup>13</sup>C Direct‐Detection Biomolecular NMR Spectroscopy in Living Cells

Bertini, Ivano; Felli, Isabella C.; Gonnelli, Leonardo; Kumar, M V Vasantha; Pierattelli, Roberta

doi:10.1002/anie.201006636

Cited by 56 publications

(45 citation statements)

References 33 publications

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“…53–59 Recent reports, however, have disputed the longstanding notion that α-synuclein is a disordered monomer, and have instead suggested the formation of a helical tetramer under physiological conditions. 60,61 In either case, it is hoped that structural analyses of WT α-synuclein will provide insight into its function and also the onset and progression of neurodegenerative diseases.…”

Section: Historical Perspective and Biomedical Relevancementioning

confidence: 99%

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Biophysical characterization of α-synuclein and its controversial structure

Alderson

Markley

2013

Intrinsically Disordered Proteins

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α-synuclein, a presynaptic protein of poorly defined function, constitutes the main component of Parkinson disease-associated Lewy bodies. Extensive biophysical investigations have provided evidence that isolated α-synuclein is an intrinsically disordered protein (IDP) in vitro. Subsequently serving as a model IDP in numerous studies, α-synuclein has aided in the development of many technologies used to characterize IDPs and arguably represents the most thoroughly analyzed IDP to date. Recent reports, however, have challenged the disordered nature of α-synuclein inside cells and have instead proposed a physiologically relevant helical tetramer. Despite α-synuclein’s rich biophysical history, a single coherent picture has not yet emerged concerning its in vivo structure, dynamics, and physiological role(s). We present herein a review of the biophysical discoveries, developments, and models pertinent to the characterization of α-synuclein’s structure and analysis of the native tetramer controversy.

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Section: Historical Perspective and Biomedical Relevancementioning

confidence: 99%

“…11B) unambiguously confirmed that α-synuclein was a disordered monomer inside intact E. coli cells, which was consistent with previous studies. 59,120–123 …”

Section: Disorder In the Court: Order?mentioning

confidence: 99%

Biophysical characterization of α-synuclein and its controversial structure

Alderson

Markley

2013

Intrinsically Disordered Proteins

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“…Direct 13 C detection experiments are widely used to study IDPs due to the large dispersion of C′,N correlations in 2D CON spectra, and to the negligible sensitivity to solventexchange processes, which are quite pronounced in disordered regions. In addition, 13 C detection schemes were originally developed for the study of paramagnetic metalloproteins, to take advantage of the reduced sensitivity of heteronuclei to dipolar contributions and paramagnetic relaxation, which depend on the gyromagnetic ratio of the nuclear spins under observation.…”

Section: Figurementioning

confidence: 99%

NMR Characterization of Long‐Range Contacts in Intrinsically Disordered Proteins from Paramagnetic Relaxation Enhancement in ¹³C Direct‐Detection Experiments

et al. 2018

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Intrinsically disordered proteins (IDPs) carry out many biological functions. They lack a stable 3D structure and are able to adopt many different conformations in dynamic equilibrium. The interplay between local dynamics and global rearrangements is key for their function. A widely used experimental NMR spectroscopy approach to study long‐range contacts in IDPs exploits paramagnetic effects, and 1H detection experiments are generally used to determine paramagnetic relaxation enhancement (PRE) for amide protons. However, under physiological conditions, exchange broadening hampers the detection of solvent‐exposed amide protons, which reduces the content of information available. Herein, we present an experimental approach based on direct carbon detection of PRE that provides improved resolution, reduced sensitivity to exchange broadening, and complementary information derived from the use of different starting polarization sources.

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“…但显微注射通常只适合于比较大的细胞如卵母细胞(细胞直径约为 1 mm)等 [57] . 虽然 In-cell NMR 近年来有较大的发展, 但在所有报道的液体 in-cell NMR 研究中, 其中目的蛋白质的 NMR 线宽 kDa 的蛋白质, 绝大部分蛋白质, 包括很多分子量同样小于 10 kDa 的蛋白质不能用传统的核磁共振方法获得高分辨谱 [49,50,58,59] . In-cell NMR 目前还处于起步阶段, 需要大力发展新的标记方法, 脉冲序列和快速采样及信号处理方法以克服灵敏度低和背景信号干扰等问题.…”

Section: In-cell Nmrunclassified

Recent Progress in the Nuclear Magnetic Resonance Applications in Analytical Chemistry

Wang¹,

Ma²,

Liu³

2012

Acta Chim. Sinica

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Development of new and effective methods for measuring chemical composition, molecular structures, interactions and dynamics is one of the major issues of analytical chemistry. Spectral analysis (spectroscopy, mass spectrometry and nuclear magnetic resonance) is the most commonly used analytical tool to address these issues. Nuclear magnetic resonance is capable to determine structure for small molecules, macromolecules and complicated biological systems, and it is considered as the most powerful tool in analytical chemistry. This paper reviewed recent progress of nuclear magnetic resonance in biological macromolecules system, complex system and the hyphenated method applications in analytical chemistry. In the first part, we gave a brief introduction of nuclear magnetic resonance technology and its applications in analytical chemistry field. The detailed application descriptions of nuclear magnetic resonance technology have been summarized from part two to part four. In the second part, we summarized the applications of nuclear magnetic resonance technology in biological macromolecules system, including the main nuclear magnetic resonance technology development in three dimensional protein structural analysis field and its applications; the related methods and applications in the dynamic study of protein complex, the in-cell nuclear magnetic resonance labeling methods development history and its applications, and also the methods of nuclear magnetic resonance technologies in studying the interactions of protein and drugs. In the third part, the qualitative and quantitative analysis of nuclear magnetic resonance technologies in complex systems has been summarized, including the applications in the metabolomics and the applications in the field of food quality and safety. In the fourth part, we briefly introduced the joint applications of magnetic resonance technologies and other separation methods such as chromatography and spectroscopic ways. The conclusions of nuclear magnetic resonance technology applications and the suggestions of future developing directions were stated in the last part.

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¹³C Direct‐Detection Biomolecular NMR Spectroscopy in Living Cells

Cited by 56 publications

References 33 publications

Biophysical characterization of α-synuclein and its controversial structure

Biophysical characterization of α-synuclein and its controversial structure

NMR Characterization of Long‐Range Contacts in Intrinsically Disordered Proteins from Paramagnetic Relaxation Enhancement in ¹³C Direct‐Detection Experiments

Recent Progress in the Nuclear Magnetic Resonance Applications in Analytical Chemistry

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13C Direct‐Detection Biomolecular NMR Spectroscopy in Living Cells

Cited by 56 publications

References 33 publications

Biophysical characterization of α-synuclein and its controversial structure

Biophysical characterization of α-synuclein and its controversial structure

NMR Characterization of Long‐Range Contacts in Intrinsically Disordered Proteins from Paramagnetic Relaxation Enhancement in 13C Direct‐Detection Experiments

Recent Progress in the Nuclear Magnetic Resonance Applications in Analytical Chemistry

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¹³C Direct‐Detection Biomolecular NMR Spectroscopy in Living Cells

NMR Characterization of Long‐Range Contacts in Intrinsically Disordered Proteins from Paramagnetic Relaxation Enhancement in ¹³C Direct‐Detection Experiments