Localising nuclear spins by pseudocontact shifts from a single tagging site.

Ligating a protein at a specific site with a tag molecule containing a paramagnetic metal ion provides a versatile way of generating pseudocontact shifts (PCSs) in nuclear magnetic resonance (NMR) spectra. PCSs can be observed for nuclear spins far from the tagging site, and PCSs generated from multiple tagging sites have been shown to enable highly accurate structure determinations at specific sites of interest, even when using flexible tags, provided the fitted effective magnetic susceptibility anisotropy () tensors accurately back-calculate the experimental PCSs measured in the immediate vicinity of the site of interest. The present work investigates the situation where only the local structure of a protein region or bound ligand is to be determined rather than the structure of the entire molecular system.

Paramagnetic Chemical Probes for Studying Biological Macromolecules.

Paramagnetic chemical probes have been used in electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) spectroscopy for more than four decades. Recent years witnessed a great increase in the variety of probes for the study of biological macromolecules (proteins, nucleic acids, and oligosaccharides). This Review aims to provide a comprehensive overview of the existing paramagnetic chemical probes, including chemical synthetic approaches, functional properties, and selected applications.

Localising individual atoms of tryptophan side chains in the metallo--lactamase IMP-1 by pseudocontact shifts from paramagnetic lanthanoid tags at multiple sites.

The metallo--lactamase IMP-1 features a flexible loop near the active site that assumes different conformations in single crystal structures, which may assist in substrate binding and enzymatic activity. To probe the position of this loop, we labelled the tryptophan residues of IMP-1 with 7-C-indole and the protein with lanthanoid tags at three different sites. The magnetic susceptibility anisotropy () tensors were determined by measuring pseudocontact shifts (PCSs) of backbone amide protons.

Organoarsenic probes to study proteins by NMR spectroscopy.

Arsenical probes enable structural studies of proteins. We report the first organoarsenic probes for nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR) spectroscopy to study proteins in solutions. These probes can be attached to irregular loop regions.

Through-Space Scalar F-F Couplings between Fluorinated Noncanonical Amino Acids for the Detection of Specific Contacts in Proteins.

Fluorine atoms are known to display scalar F-F couplings in nuclear magnetic resonance (NMR) spectra when they are sufficiently close in space for nonbonding orbitals to overlap. We show that fluorinated noncanonical amino acids positioned in the hydrophobic core or on the surface of a protein can be linked by scalar through-space F-F () couplings even if the F spins are in the time average separated by more than the van der Waals distance. Using two different aromatic amino acids featuring CF groups, -trifluoromethyl-tyrosine and 4-trifluoromethyl-phenylalanine, we show that F-F TOCSY experiments are sufficiently sensitive to detect couplings between 2.

Paramagpy: software for fitting magnetic susceptibility tensors using paramagnetic effects measured in NMR spectra.

Paramagnetic metal ions with fast-relaxing electrons generate pseudocontact shifts (PCSs), residual dipolar couplings (RDCs), paramagnetic relaxation enhancements (PREs) and cross-correlated relaxation (CCR) in the nuclear magnetic resonance (NMR) spectra of the molecules they bind to. These effects offer long-range structural information in molecules equipped with binding sites for such metal ions. Here we present the new open-source software Paramagpy, which has been written in Python 3 with a graphic user interface.

Protein NMR Resonance Assignment without Spectral Analysis: 5D SOlid-State Automated Projection SpectroscopY (SO-APSY).

Narrow proton signals, high sensitivity, and efficient coherence transfers provided by fast magic-angle spinning at high magnetic fields make automated projection spectroscopy feasible for the solid-state NMR analysis of proteins. We present the first ultrahigh dimensional implementation of this approach, where 5D peak lists are reconstructed from a number of 2D projections for protein samples of different molecular sizes and aggregation states, which show limited dispersion of chemical shifts or inhomogeneous broadenings. The resulting datasets are particularly suitable to automated analysis and yield rapid and unbiased assignments of backbone resonances.

Accurate Electron-Nucleus Distances from Paramagnetic Relaxation Enhancements.

Measurements of paramagnetic relaxation enhancements (PREs) in H NMR spectra are an important tool to obtain long-range distance information in proteins, but quantitative interpretation is easily compromised by nonspecific intermolecular PREs. Here we show that PREs generated by lanthanides with anisotropic magnetic susceptibilities offer a route to accurate calibration-free distance measurements. As these lanthanides change H chemical shifts due to pseudocontact shifts, the relaxation rates in the paramagnetic and diamagnetic state can be measured with a single sample that simultaneously contains the protein labeled with a paramagnetic and a diamagnetic lanthanide ion.

Highly sensitive biosensors based on all-dielectric nanoresonators.

Biosensing based on nanophotonic structures has shown a great potential for cost-efficient, high-speed and compact personal medical diagnostics. While plasmonic nanosensors offer high sensitivity, their intrinsically restricted resonance quality factors and strong heating due to metal absorption impose severe limitations on real life applications. Here, we demonstrate an all-dielectric sensing platform based on silicon nanodisks with strong optically-induced magnetic resonances, which are able to detect a concentration of streptavidin of as low as 10 M (mol L) or 5 ng mL, thus pushing the current detection limit by at least two orders of magnitudes.

Refractive index sensing with Fano resonances in silicon oligomers.

We demonstrate experimentally refractive index sensing with localized Fano resonances in silicon oligomers, consisting of six disks surrounding a central one of slightly different diameter. Owing to the low absorption and narrow Fano-resonant spectral features appearing as a result of the interference of the modes of the outer and the central disks, we demonstrate refractive index sensitivity of more than 150 nm RIU with a figure of merit of 3.8.

Using Paramagnetism to Slow Down Nuclear Relaxation in Protein NMR.

Paramagnetic metal ions accelerate nuclear spin relaxation; this effect is widely used for distance measurement and called paramagnetic relaxation enhancement (PRE). Theoretical predictions established that, under special circumstances, it is also possible to achieve a reduction in nuclear relaxation rates (negative PRE). This situation would occur if the mechanism of nuclear relaxation in the diamagnetic state is counterbalanced by a paramagnetic relaxation mechanism caused by the metal ion.