Multimodal Response to Copper Binding in Superoxide Dismutase Dynamics

Bonaccorsi, Marta; Knight, Michael; Marchand, Tanguy Le; Dannatt, Hugh R. W.; Schubeis, Tobias; Salmon, Loïc; Felli, Isabella C.; Emsley, Lyndon; Pierattelli, Roberta; Pintacuda, Guido

doi:10.1021/jacs.0c09242

Cited by 18 publications

(11 citation statements)

References 63 publications

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“…Investigations of the fully mature functional Cu, Zn state of superoxide dismutase and its E, Zn-state in which the Cu site is empty have shown that the metal ion after uptake does not act as a rigidification element. Using 15 N R 1 , R 1ρ experiments, it has been shown that metal binding acts rather as a switch that redistributes motional processes on different time scales and induces a coupling of the dynamics of the histidine side chains to remote key backbone elements of the protein . The SAIL approach allows selectivity of isotopically labeled aromatic rings in proteins .…”

Section: Quantification Of Dynamicsmentioning

confidence: 99%

Deuteration for High-Resolution Detection of Protons in Protein Magic Angle Spinning (MAS) Solid-State NMR

Reif

2021

Chem. Rev.

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Proton detection developed in the last 20 years as the method of choice to study biomolecules in the solid state. In perdeuterated proteins, proton dipolar interactions are strongly attenuated, which allows yielding of high-resolution proton spectra. Perdeuteration and backsubstitution of exchangeable protons is essential if samples are rotated with MAS rotation frequencies below 60 kHz. Protonated samples can be investigated directly without spin dilution using proton detection methods in case the MAS frequency exceeds 110 kHz. This review summarizes labeling strategies and the spectroscopic methods to perform experiments that yield assignments, quantitative information on structure, and dynamics using perdeuterated samples. Techniques for solvent suppression, H/D exchange, and deuterium spectroscopy are discussed. Finally, experimental and theoretical results that allow estimation of the sensitivity of proton detected experiments as a function of the MAS frequency and the external B 0 field in a perdeuterated environment are compiled.

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Section: Quantification Of Dynamicsmentioning

confidence: 99%

Deuteration for High-Resolution Detection of Protons in Protein Magic Angle Spinning (MAS) Solid-State NMR

Reif

2021

Chem. Rev.

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“…We additionally present a comparison of the results obtained with chemical shifts previously obtained on deuterated MBP in solution (Evenäs et al 2001). Considering the recent findings of previously unobserved dynamics in other, supposedly rigid, crystalline proteins (Rovó et al 2019;Bonaccorsi et al 2020), we additionally anticipate the possibility of new insight on internal motions which escaped previous determinations by X-ray and solution NMR.…”

Section: Introductionmentioning

confidence: 87%

Backbone assignment of crystalline E. coli maltose binding protein

2021

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The E.coli maltose binding protein (MBP) is a 42.5 kDa molecule widely employed in many biotechnology applications. Because of its molecular size, it has become the main model system for the development of solution NMR methods adapted to large biomolecular targets. Here, we report virtually complete (~90%) backbone resonance assignments obtained on a microcrystalline sample of MBP with 1 H-detected solidstate NMR at fast (>100 kHz) magic-angle spinning. We additionally present the detailed description of the methodology employed for the preparation of the sample and the acquisition and analysis of the NMR spectra. The chemical shifts, obtained with a single uniformly 15 N, 13 C-labelled and fully-protonated sample and about two weeks on a 800 MHz NMR spectrometer, have been deposited to the BMRB under the accession number 50089.

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“…Conversely the investigation of motions in a window from ps to ms revealed a multimodal response, with the metal acting as a control element and redistributing dynamic processes over multiple timescales. Moreover, a coupling was observed between motions of side chains coordinating the copper in the active site and those of distant backbone elements of the protein which are pivotal for SOD stability and efficient enzymatic activity [33]. The multitimescale dynamics unveiled in these studies by solid-state NMR are particularly difficult to observe using other methods [34], and in the specific case of SOD had escaped previous determinations by solution NMR.…”

Section: Introductionmentioning

confidence: 95%