[NMR paper] Rapid Quantitative Measurements of Paramagnetic Relaxation Enhancements in Cu(II)-Tagged Proteins by Proton-Detected Solid-State NMR Spectroscopy.

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Rapid Quantitative Measurements of Paramagnetic Relaxation Enhancements in Cu(II)-Tagged Proteins by Proton-Detected Solid-State NMR Spectroscopy.

Related Articles Rapid Quantitative Measurements of Paramagnetic Relaxation Enhancements in Cu(II)-Tagged Proteins by Proton-Detected Solid-State NMR Spectroscopy.

J Phys Chem Lett. 2017 Nov 17;:

Authors: Mukhopadhyay D, Nadaud PS, Shannon MD, Jaroniec CP

Abstract
We demonstrate rapid quantitative measurements of site-resolved paramagnetic relaxation enhancements (PREs), which are a source of valuable structural restraints corresponding to electron-nucleus distances in the ~10-20 Å regime, in solid-state nuclear magnetic resonance (NMR) spectra of proteins containing covalent Cu(2+)-binding tags. Specifically, using protein GB1 K28C-EDTA-Cu(2+) mutant as a model, we show the determination of backbone amide (15)N longitudinal and (1)H transverse PREs within a few hours of experiment time based on proton-detected 2D or 3D correlation spectra recorded with magic-angle spinning frequencies >= ~60 kHz for samples containing ~10-50 nanomoles of (2)H,(13)C,(15)N-labeled protein back-exchanged in H2O. Additionally, we show that the electron relaxation time for the Cu(2+) center, needed to convert PREs into distances, can be estimated directly from the experimental data. Altogether, these results are important for establishing solid-state NMR based on paramagnetic-tagging as a routine tool for structure determination of natively diamagnetic proteins.


PMID: 29148785 [PubMed - as supplied by publisher]



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