Dataset Information

Sub-Micromolar Pulse Dipolar EPR Spectroscopy Reveals Increasing CuII -labelling of Double-Histidine Motifs with Lower Temperature.

ABSTRACT: Electron paramagnetic resonance (EPR) distance measurements are making increasingly important contributions to the studies of biomolecules by providing highly accurate geometric constraints. Combining double-histidine motifs with CuII spin labels can further increase the precision of distance measurements. It is also useful for proteins containing essential cysteines that can interfere with thiol-specific labelling. However, the non-covalent CuII coordination approach is vulnerable to low binding-affinity. Herein, dissociation constants (KD ) are investigated directly from the modulation depths of relaxation-induced dipolar modulation enhancement (RIDME) EPR experiments. This reveals low- to sub-?m CuII KD s under EPR distance measurement conditions at cryogenic temperatures. We show the feasibility of exploiting the double-histidine motif for EPR applications even at sub-?m protein concentrations in orthogonally labelled CuII -nitroxide systems using a commercial Q-band EPR instrument.

SUBMITTER: Wort JL

PROVIDER: S-EPMC6771633 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

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Sub-Micromolar Pulse Dipolar EPR Spectroscopy Reveals Increasing CuII -labelling of Double-Histidine Motifs with Lower Temperature.

Wort Joshua L JL Ackermann Katrin K Giannoulis Angeliki A Stewart Alan J AJ Norman David G DG Bode Bela E BE

Angewandte Chemie (International ed. in English) 20190718 34

Electron paramagnetic resonance (EPR) distance measurements are making increasingly important contributions to the studies of biomolecules by providing highly accurate geometric constraints. Combining double-histidine motifs with CuII spin labels can further increase the precision of distance measurements. It is also useful for proteins containing essential cysteines that can interfere with thiol-specific labelling. However, the non-covalent CuII coordination approach is vu ...[more]

PMID: 31218813

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Project description:BackgroundThere are a range of interventions designed to promote healthier food choices in full-service restaurants. However, it is unclear how these interventions affect dietary choices in people of lower and higher socioeconomic position (SEP).MethodsA total of 2091 US participants recruited online completed Study 1 (n = 1001) and Study 2 (n = 1090). Recruitment was stratified by participant highest education level, resulting in higher SEP and lower SEP groups. In a between-subjects design, participants made hypothetical food choices (main dish, plus optional sides and desserts) from six restaurants menus in the absence vs. presence of menu energy labelling and from menus with baseline (10%) vs. increased availability (50%) of lower energy main dishes. Data were collected and analysed in 2019. Two studies were conducted in order to examine replicability and generalisability of findings across different restaurant menu types.ResultsAcross both studies, increasing the availability of lower energy main menu options decreased the average energy content of the ordered main dish (- 129 kcal, 95% CI [- 139; - 119]) and total energy ordered (- 117 kcal, 95% CI [- 138; - 95]) in both higher and lower SEP participants. Energy labelling significantly reduced the energy content of ordered main dishes in higher SEP participants (- 41 kcal, 95% CI [- 54; - 29]), but not lower SEP participants (- 5 kcal, 95% CI [- 22; 11]). However, energy labelling reduced total energy ordered (- 83 kcal, 95% CI [- 105; - 60]) irrespective of SEP.ConclusionsIn two virtual experiments, increasing the availability of lower energy restaurant main menu options impacted on main menu dish choice and decreased total energy ordered irrespective of SEP. Energy labelling had a less pronounced effect on total energy ordered and had a larger impact on the energy content of main menu dish choice in higher as opposed to lower SEP participants.Trial registrationClinicaltrials.gov NCT04336540 retrospectively registered (7 April, 2020).

Dataset Information

Sub-Micromolar Pulse Dipolar EPR Spectroscopy Reveals Increasing CuII -labelling of Double-Histidine Motifs with Lower Temperature.

Publications

Sub-Micromolar Pulse Dipolar EPR Spectroscopy Reveals Increasing Cu<sup>II</sup> -labelling of Double-Histidine Motifs with Lower Temperature.

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