Supramolecular Approach to Electron Paramagnetic Resonance Distance Measurement of Spin-Labeled Proteins
| Autor: | Gareth R. Eaton, Zhimin Yang, Gunnar Jeschke, Maren Pink, Sandra S. Eaton, Richard A. Stein, Hassane S. Mchaourab, Andrzej Rajca, Thacien Ngendahimana, Suchada Rajca |
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| Rok vydání: | 2020 |
| Předmět: |
Materials science
Electron Spin Resonance Spectroscopy Temperature Supramolecular chemistry Proteins Resonance macromolecular substances Article Surfaces Coatings and Films law.invention chemistry.chemical_compound Nuclear magnetic resonance Molecular recognition Distance measurement chemistry law Materials Chemistry Iodoacetamide Muramidase Spin Labels Physical and Theoretical Chemistry Electron paramagnetic resonance Spin labeled |
| Zdroj: | J Phys Chem B |
| ISSN: | 1520-5207 1520-6106 |
| DOI: | 10.1021/acs.jpcb.0c00743 |
| Popis: | We demonstrate a host-guest molecular recognition approach to advance double electron-electron resonance (DEER) distance measurements of spin-labeled proteins. We synthesized an iodoacetamide (IA) derivative of 2,6-diazaadamantane nitroxide (DZD) spin label that could be doubly incorporated into T4 Lysozyme (T4L) by site-directed spin labeling (SDSL) with efficiency up to 50% per cysteine. The rigidity of the fused ring structure and absence of mobile methyl groups increase the spin echo dephasing time (T(m)) at temperatures above 80 K. This enables DEER measurements of distances >4 nm in DZD labeled-T4L in glycerol/water at temperatures up to 150 K, with increased sensitivity compared to common spin label such as MTSL. Addition of β-cyclodextrin (β-CD) reduces the rotational correlation time of the label, slightly increases T(m), and most importantly, narrows (and slightly lengthens) the inter-spin distance distributions. The distance distributions are in good agreement with simulated distance distributions obtained by rotamer libraries. These results provide a foundation for developing supramolecular recognition to facilitate long-distance DEER measurements at near physiological temperatures. |
| Databáze: | OpenAIRE |
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