Encoded loop-lanthanide-binding tags for long-range distance measurements in proteins by NMR and EPR spectroscopy

Journal of Biomolecular NMR
Dominic BarthelmesHarald Schwalbe

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 (1)H, (15)N-pseudocontact shifts (PCSs) by NMR spectroscopy. We determined the Δχ-tensors associated with each Tm(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(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.

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Citations

Dec 2, 2015·The Journal of Physical Chemistry Letters·Elwy H AbdelkaderDaniella Goldfarb
Dec 10, 2016·Journal of the American Chemical Society·Diane DaubeBjörn Corzilius
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Oct 7, 2016·Physical Chemistry Chemical Physics : PCCP·Katharina KellerMaxim Yulikov
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Sep 6, 2018·Physical Chemistry Chemical Physics : PCCP·Mithun C MahawaththaGottfried Otting
Mar 13, 2019·Physical Chemistry Chemical Physics : PCCP·Angeliki GiannoulisDaniella Goldfarb
May 24, 2017·Journal of Biomolecular NMR·Dominic BarthelmesHarald Schwalbe
Aug 3, 2017·The Journal of Chemical Physics·Nurit ManukovskyDaniella Goldfarb
Jan 19, 2019·International Journal of Molecular Sciences·Theresa BraunDaniel Summerer
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