ArticleJournal of the American Chemical Society2011
Engineering encodable lanthanide-binding tags into loop regions of proteins.
Article in Journal of the American Chemical Society, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers.
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Who cites it
46 citing papers in PubMed.
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- Solution Structure of a Lanthanide-binding DNA Aptamer Determined Using High Quality pseudocontact shift restraints.Chemistry (Weinheim an der Bergstrasse, Germany) · 2022Article
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- Paramagnetic Chemical Probes for Studying Biological Macromolecules.Chemical reviews · 2022Review
- An automated iterative approach for protein structure refinement using pseudocontact shifts.Journal of biomolecular NMR · 2021Article
- Conformational ensemble of a multidomain protein explored by GdBiophysical journal · 2021Article
- Phosphoserine for the generation of lanthanide-binding sites on proteins for paramagnetic nuclear magnetic resonance spectroscopy.Magnetic resonance (Gottingen, Germany) · 2021Article
- Designed Metal-ATCUN Derivatives: Redox- and Non-redox-Based Applications Relevant for Chemistry, Biology, and Medicine.iScience · 2020Review
- Lanthanide Tagging of Oligonucleotides to Nucleobase for Paramagnetic NMR.Chembiochem : a European journal of chemical biology · 2020Article
- The copper(II)-binding tripeptide GHK, a valuable crystallization and phasing tag for macromolecular crystallography.Acta crystallographica. Section D, Structural biology · 2020Article
- Paramagnetic NMR in drug discovery.Journal of biomolecular NMR · 2020Review
- A Double-Armed, Hydrophilic Transition Metal Complex as a Paramagnetic NMR Probe.Angewandte Chemie (International ed. in English) · 2019Review
- Theory and practice of using solvent paramagnetic relaxation enhancement to characterize protein conformational dynamics.Methods (San Diego, Calif.) · 2018Article
- Structural Characterization of a Heparan Sulfate Pentamer Interacting with LAR-Ig1-2.Biochemistry · 2018Article
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8 authors.
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Abstract
Lanthanide-binding tags (LBTs) are valuable tools for investigation of protein structure, function, and dynamics by NMR spectroscopy, X-ray crystallography, and luminescence studies. We have inserted LBTs into three different loop positions (denoted L, R, and S) of the model protein interleukin-1β (IL1β) and varied the length of the spacer between the LBT and the protein (denoted 1−3). Luminescence studies demonstrate that all nine constructs bind Tb3+ tightly in the low nanomolar range. No significant change in the fusion protein occurs from insertion of the LBT, as shown by two X-ray crystallographic structures of the IL1β-S1 and IL1β-L3 constructs and for the remaining constructs by comparing the 1H−15N heteronuclear single-quantum coherence NMR spectra with that of the wild-type IL1β. Additionally, binding of LBT-loop IL1β proteins to their native binding partner in vitro remains unaltered. X-ray crystallographic phasing was successful using only the signal from the bound lanthanide. Large residual dipolar couplings (RDCs) could be determined by NMR spectroscopy for all LBT-loop constructs and revealed that the LBT-2 series were rigidly incorporated into the interleukin-1β structure. The paramagnetic NMR spectra of loop-LBT mutant IL1β-R2 were assigned and the Δχ tensor components were calculated on the basis of RDCs and pseudocontact shifts. A structural model of the IL1β-R2 construct was calculated using the paramagnetic restraints. The current data provide support that encodable LBTs serve as versatile biophysical tags when inserted into loop regions of proteins of known structure or predicted via homology modeling.
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