Encoded loop-lanthanide-binding tags for long-range distance measurements in proteins by NMR and EPR spectroscopy
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10858_2015_Article_9984.pdf
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Author(s)
Barthelmes, Dominic
Gränz, Markus
Barthelmes, Katja
Allen, Karen N.
Imperiali, Barbara
Prisner, Thomas
Schwalbe, Harald
Date Issued
September 2015
Journal
Journal of Biomolecular NMR
Publisher
Springer Netherlands
Citation
Barthelmes, Dominic, Markus Gränz, Katja Barthelmes, Karen N. Allen, Barbara Imperiali, Thomas Prisner, and Harald Schwalbe. “Encoded Loop-Lanthanide-Binding Tags for Long-Range Distance Measurements in Proteins by NMR and EPR Spectroscopy.” J Biomol NMR 63, no. 3 (September 4, 2015): 275–282.
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Author's final manuscript
Abstract
We recently engineered encodable lanthanide binding tags (LBTs) into proteins and demonstrated their applicability in Nuclear Magnetic Resonance (NMR) spectroscopy, X-ray crystallography and luminescence studies. Here, we engineered two-loop-LBTs into the model protein interleukin-1β (IL1β) and measured [superscript 1]H, [superscript 15]N-pseudocontact shifts (PCSs) by NMR spectroscopy. We determined the Δχ-tensors associated with each Tm[superscript 3+]-loaded loop-LBT and show that the experimental PCSs yield structural information at the interface between the two metal ion centers at atomic resolution. Such information is very valuable for the determination of the sites of interfaces in protein–protein-complexes. Combining the experimental PCSs of the two-loop-LBT construct IL1β-S2R2 and the respective single-loop-LBT constructs IL1β-S2, IL1β-R2 we additionally determined the distance between the metal ion centers. Further, we explore the use of two-loop LBTs loaded with Gd[superscript 3+] as a novel tool for distance determination by Electron Paramagnetic Resonance spectroscopy and show the NMR-derived distances to be remarkably consistent with distances derived from Pulsed Electron–Electron Dipolar Resonance.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Chemistry
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DOI of Published Version
https://doi.org/10.1007/s10858-015-9984-x
