Authors: Luchinat E, Barbieri L, Cremonini M, Banci L
Abstract
In-cell NMR spectroscopy provides precious structural and functional information on biological macromolecules in their native cellular environment at atomic resolution. However, the intrinsic low sensitivity of NMR imposes a big limitation in the applicability of the methodology. In this respect, the recently developed commercial 1.2*GHz NMR spectrometer is expected to introduce significant benefits. However, cell samples may suffer from detrimental effects at ultrahigh fields, that must be carefully evaluated. Here we show the first in-cell NMR spectra recorded at 1.2*GHz on human cells, and we compare resolution and sensitivity against those obtained at 900 and 950*MHz. To evaluate the effects of different spin relaxation rates, SOFAST-HMQC and BEST-TROSY spectra were recorded on intracellular ?-synuclein and carbonic anhydrase. Major improvements are observed at 1.2*GHz when analyzing unfolded proteins, such as ?-synuclein, while the TROSY scheme improves the resolution for both globular and unfolded proteins.
PMID: 33580357 [PubMed - as supplied by publisher]
Protein in-cell NMR spectroscopy at 1.2Â*GHz
Protein in-cell NMR spectroscopy at 1.2Â*GHz
Abstract
In-cell NMR spectroscopy provides precious structural and functional information on biological macromolecules in their native cellular environment at atomic resolution. However, the intrinsic low sensitivity of NMR imposes a big limitation in the applicability of the methodology. In this respect, the recently developed commercial 1.2Â*GHz NMR spectrometer is expected to introduce significant benefits. However, cell samples may suffer from detrimental effects at ultrahigh fields, that must be...
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15N isotopic labelling for in-cell protein studies by NMR spectroscopy and single-cell IR synchrotron radiation FTIR microscopy: a correlative study.
15N isotopic labelling for in-cell protein studies by NMR spectroscopy and single-cell IR synchrotron radiation FTIR microscopy: a correlative study.
Analyst. 2018 Feb 06;:
Authors: Mitri E, Barbieri L, Vaccari L, Luchinat E
Abstract
The ultimate goal of modern structural biology is to probe protein structures and dynamics in their physiological microenvironment. In-cell NMR...
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Abstract
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Angew Chem Int Ed Engl. 2015 Mar 5;
Authors: Wen H, An YJ, Xu WJ, Kang KW, Park S
Abstract
Altered metabolism is a critical part of...
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Chembiochem. 2014 Apr 1;
Authors: Majumder S, Demott CM, Burz DS, Shekhtman A
Abstract
Distinct differences between how model proteins interact in-cell and in vitro suggest that the cytosol might have a profound effect in modulating protein-protein and/or protein-ligand interactions that are...
Protein in-cell NMR spectroscopy at 1.2*GHz.
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