A novel, rationally designed lanthanoid chelating tag delivers large paramagnetic structural restraints for biomolecular NMR

Joss, Daniel; Winter, Florine; Häußinger, Daniel

doi:10.1039/d0cc04337k

Cited by 11 publications

(20 citation statements)

References 36 publications

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“…287 Recent work introduced a 3-nitro-substituted 2-(methylsulfonyl)pyridine moiety as attachment group to design a new DOTA-M8 based probe named M7-Nitro (Figure 9, 93). 288 This probe possesses high protein ligation rates (more than 95% ligation in 30 minutes at room temperature) and a very large Δχ tensor (Δχ ax = 94.3 × 10 −32 m 3 ) was measured with Dy(III)-M7-Nitro labeled ubiquitin S57C. The authors attributed the improvements in ligation to the methylsulfone leaving group in the ortho position and the presence of the nitro substituent in the meta position.…”

Section: Chemicalmentioning

confidence: 99%

Paramagnetic Chemical Probes for Studying Biological Macromolecules

et al. 2022

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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. Recent developments have seen, in particular, a rapid expansion of the range of lanthanoid probes with anisotropic magnetic susceptibilities for the generation of structural restraints based on residual dipolar couplings and pseudocontact shifts in solution and solid state NMR spectroscopy, mostly for protein studies. Also many new isotropic paramagnetic probes, suitable for NMR measurements of paramagnetic relaxation enhancements, as well as EPR spectroscopic studies (in particular double resonance techniques) have been developed and employed to investigate biological macromolecules. Notwithstanding the large number of reported probes, only few have found broad application and further development of probes for dedicated applications is foreseen.

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Section: Chemicalmentioning

confidence: 99%

Paramagnetic Chemical Probes for Studying Biological Macromolecules

et al. 2022

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“…The anisotropy values were determined from the shifts of the ligand nuclei in the case of the DOTA, 71 DOTA-M8, 72 DOTA-M7FPy 72 and tris(dipicolinato) 73,74 complexes, and from the nuclear shifts of the attached proteins in the case of the tags CLaNP-5, 28,75 BrPSPy-DO3M(S)A 76 and Ln-M7-Nitro. 77 In the case of the proteins with paramagnetic tags, the shifts may be reduced to some extent by a non-completely rigid attachment of the tag.…”

Section: Magnetic Susceptibility Anisotropy and Electron Relaxation R...mentioning

confidence: 99%

The evolution of paramagnetic NMR as a tool in structural biology

Ravera

Gigli

Fiorucci

et al. 2022

Phys. Chem. Chem. Phys.

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“…[ 48 , 49 , 50 ] vi) Synthesis of the lanthanide tag should be straightforward and affordable. Among the many tags published to date, the P4T‐DOTA and Ln‐M7‐Nitro tags recently published by Häussinger and co‐workers[ 51 , 52 ] (Figure 1 ) fulfil these criteria, except that their synthesis is challenging.…”

Section: Introductionmentioning

confidence: 99%

A Chiral Lanthanide Tag for Stable and Rigid Attachment to Single Cysteine Residues in Proteins for NMR, EPR and Time‐Resolved Luminescence Studies

Herath

Breen

Hewitt

et al. 2021

Chemistry A European J

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A lanthanide-binding tag site-specifically attached to a protein presents a tool to probe the protein by multiple spectroscopic techniques, including nuclear magnetic resonance, electron paramagnetic resonance and time-resolved luminescence spectroscopy. Here a new stable chiral Ln III tag, referred to as C12, is presented for spontaneous and quantitative reaction with a cysteine residue to generate a stable thioether bond. The synthetic protocol of the tag is relatively straightforward, and the tag is stable for storage and shipping. It displays greatly enhanced reactivity towards selenocysteine, opening a route towards selective tagging of selenocysteine in proteins containing cysteine residues.Loaded with Tb III or Tm III ions, the C12 tag readily generates pseudocontact shifts (PCS) in protein NMR spectra. It produces a relatively rigid tether between lanthanide and protein, which is beneficial for interpretation of the PCSs by single magnetic susceptibility anisotropy tensors, and it is suitable for measuring distance distributions in double electron-electron resonance experiments. Upon reaction with cysteine or other thiol compounds, the Tb III complex exhibits a 100-fold enhancement in luminescence quantum yield, affording a highly sensitive turn-on luminescence probe for time-resolved FRET assays and enzyme reaction monitoring.

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A novel, rationally designed lanthanoid chelating tag delivers large paramagnetic structural restraints for biomolecular NMR

Abstract: Herein, a novel and rationally designed ortho-substituted pyridine activator is reported that reacts rapidly and selectively with cysteine thiols. It forms reduction-stable conjugates and induces large pseudocontact shifts, residual dipolar...

Cited by 11 publications

References 36 publications

Paramagnetic Chemical Probes for Studying Biological Macromolecules

Paramagnetic Chemical Probes for Studying Biological Macromolecules

The evolution of paramagnetic NMR as a tool in structural biology

A Chiral Lanthanide Tag for Stable and Rigid Attachment to Single Cysteine Residues in Proteins for NMR, EPR and Time‐Resolved Luminescence Studies

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