Lanthanide labeling offers fast NMR approach to 3D structure determinations of protein-protein complexes

Journal of the American Chemical Society
Guido PintacudaGottfried Otting

Abstract

A novel nuclear magnetic resonance (NMR) strategy based on labeling with lanthanides achieves rapid determinations of accurate three-dimensional (3D) structures of protein-protein complexes. The method employs pseudocontact shifts (PCS) induced by a site-specifically bound lanthanide ion to anchor the coordinate system of the magnetic susceptibility tensor in the molecular frames of the two molecules. Simple superposition of the tensors detected in the two protein molecules brings them together in a 3D model of the protein-protein complex. The method is demonstrated with the 30 kDa complex between two subunits of Escherichia coli polymerase III, comprising the N-terminal domain of the exonuclease subunit epsilon and the subunit theta. The 3D structures of the individual molecules were docked based on a limited number of PCS observed in 2D 15N-heteronuclear single quantum coherence spectra. Degeneracies in the mutual orientation of the protein structures were resolved by the use of two different lanthanide ions, Dy3+ and Er3+.

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Citations

Nov 11, 2006·Journal of Biomolecular NMR·Michael JohnGottfried Otting
May 10, 2007·Journal of Biomolecular NMR·Lianmei FengJames H Prestegard
Aug 9, 2008·Journal of Biomolecular NMR·Gottfried Otting
Jun 17, 2009·Journal of Biomolecular NMR·Xun-Cheng Su, Gottfried Otting
Mar 20, 2010·Journal of Biomolecular NMR·Tomohide SaioFuyuhiko Inagaki
Dec 31, 2010·Journal of Biomolecular NMR·Xingsheng WangFang Tian
Jun 1, 2011·Journal of Biomolecular NMR·Christophe Schmitz, Alexandre M J J Bonvin
Dec 25, 2012·Journal of Biomolecular NMR·Hiromasa YagiGottfried Otting
Dec 25, 2010·Journal of the American Chemical Society·Katja BarthelmesHarald Schwalbe
May 29, 2010·Microbiology and Molecular Biology Reviews : MMBR·Andrew RobinsonNicholas E Dixon
May 14, 2010·Annual Review of Biophysics·Gottfried Otting
Jun 14, 2014·Progress in Nuclear Magnetic Resonance Spectroscopy·Christoph GöblMichael Sattler
Jan 31, 2012·Journal of Molecular Biology·Christophe SchmitzThomas Huber
Jul 29, 2009·Biochimica Et Biophysica Acta·Daniela BressaninAlejandro Hochkoeppler
Aug 21, 2015·Journal of the American Chemical Society·Ulrika BrathMáté Erdélyi
Nov 7, 2006·Journal of Magnetic Resonance·Junji IwaharaG Marius Clore
Oct 17, 2009·Proteomics·Mitchell R O'ConnellJoel P Mackay
Mar 15, 2016·Journal of the American Chemical Society·Wan-Na ChenGottfried Otting
Nov 20, 2015·Scientific Reports·Tomohide SaioFuyuhiko Inagaki
Apr 12, 2014·Current Opinion in Structural Biology·Mathias A S Hass, Marcellus Ubbink
Jan 22, 2015·Journal of Biomolecular NMR·Duncan J CrickDaniel Nietlispach
Nov 25, 2014·Journal of Biomolecular NMR·Mauro RinaldelliClaudio Luchinat
Jan 10, 2018·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Walter BeckerKlaus Zangger
Oct 9, 2014·Angewandte Chemie·Aldo R Camacho-ZarcoMarkus Zweckstetter
Oct 17, 2018·Future Medicinal Chemistry·Felix Torres, Julien Orts
Nov 23, 2006·Chemistry : a European Journal·Monica D VlasieMarcellus Ubbink
Oct 3, 2007·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Michael John, Gottfried Otting
Jul 17, 2008·Dalton Transactions : an International Journal of Inorganic Chemistry·Ivano BertiniRoberta Pierattelli

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