Pulsed electron-electron double-resonance determination of spin-label distances and orientations on the tetrameric potassium ion channel KcsA.

Journal of the American Chemical Society
Burkhard EndewardThomas F Prisner

Abstract

Pulsed electron-electron double-resonance (PELDOR) measurements are presented from the potassium ion channel KcsA both solubilized in detergent and reconstituted in lipids. Site-directed spin-labeling using (1-oxyl-2,2,5,5-tetramethyl-3-pyrrolin-3-yl)methyl methanethiosulfonate was performed with a R64C mutant of the protein. The orientations of the spin-labels in the tetramer were determined by PELDOR experiments performed at two magnetic field strengths (0.3 T/X-band and 1.2 T/Q-band) and variable probe frequency. Quantitative simulation of the PELDOR data supports a strongly restricted nitroxide, oriented at an angle of 65 degrees relative to the central channel axis. In general, poorer quality PELDOR data were obtained from membrane-reconstituted preparations compared to soluble proteins or detergent-solubilized samples. One reason for this is the reduced transverse spin relaxation time T(2) of nitroxides due to crowding of tetramers within the membrane that occurs even at low protein to lipid ratios. This reduced T(2) can be overcome by reconstituting mixtures of unlabeled and labeled proteins, yielding high-quality PELDOR data. Identical PELDOR oscillation frequencies and their dependencies on the probe frequency were obs...Continue Reading

References

Sep 25, 1999·The Journal of General Physiology·L HeginbothamC Miller
Jan 29, 2000·Journal of Magnetic Resonance·M PannierH W Spiess
Aug 31, 2000·Nature Structural Biology·W L HubbellC Altenbach
Sep 27, 2002·Journal of Magnetic Resonance·Axel WeberThomas F Prisner
Jan 15, 2005·Journal of Magnetic Resonance·Yun-Wei ChiangJack H Freed
Mar 15, 2006·Nature Structural & Molecular Biology·Julio F Cordero-MoralesEduardo Perozo
Aug 30, 2006·Proceedings of the National Academy of Sciences of the United States of America·V P DenysenkovM Bennati
Oct 20, 2006·Angewandte Chemie·Adelheid GodtGunnar Jeschke
Dec 26, 2006·Journal of Magnetic Resonance·Ye PolyhachG Jeschke
Jun 15, 2007·Quarterly Reviews of Biophysics·Olav Schiemann, Thomas F Prisner
Dec 22, 2007·Protein Science : a Publication of the Protein Society·Zhefeng GuoWayne L Hubbell
Feb 19, 2009·The Journal of Chemical Physics·A MarkoT Prisner

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Citations

Oct 29, 2013·Progress in Nuclear Magnetic Resonance Spectroscopy·Klaus MöbiusAnton Savitsky
Nov 17, 2012·Chemical Reviews·Joseph W F RobertsonSoojay Banerjee
Jan 11, 2012·Journal of the American Chemical Society·Xiaojun ZhangPeter Z Qin
Apr 2, 2010·Physical Chemistry Chemical Physics : PCCP·Jan HoffmannBernd Brutschy
May 10, 2011·Physical Chemistry Chemical Physics : PCCP·Michal Gordon-GrossmanDaniella Goldfarb
Jul 7, 2011·Molecular BioSystems·N A KuznetsovYu D Tsvetkov
Jun 30, 2012·Physical Chemistry Chemical Physics : PCCP·Maxim YulikovGunnar Jeschke
Jul 4, 2012·Physical Chemistry Chemical Physics : PCCP·Yevhen PolyhachGunnar Jeschke
Mar 15, 2013·Physical Chemistry Chemical Physics : PCCP·Tona von HagensGunnar Jeschke
Jan 17, 2014·Physical Chemistry Chemical Physics : PCCP·Daniella Goldfarb
Sep 27, 2012·Proceedings of the National Academy of Sciences of the United States of America·Christos PliotasJames H Naismith
Mar 13, 2012·Annual Review of Physical Chemistry·Gunnar Jeschke
Jun 24, 2014·Molecular Physics·Angeliki GiannoulisBela E Bode
Mar 27, 2012·Applied Magnetic Resonance·Gregor HageluekenOlav Schiemann
May 15, 2012·Journal of Magnetic Resonance·Suzanne BrandonEric J Hustedt
Mar 6, 2012·Journal of Magnetic Resonance·Gunnar W ReginssonOlav Schiemann
Jan 31, 2012·Journal of Magnetic Resonance·Christoph AbéHeinz-Jürgen Steinhoff
Jan 27, 2011·Biochemical Society Transactions·Gunnar W Reginsson, Olav Schiemann
Feb 26, 2011·The Biochemical Journal·Gunnar W Reginsson, Olav Schiemann
Sep 2, 2010·Journal of Magnetic Resonance·Richard WardDavid G Norman
Feb 24, 2015·Journal of Magnetic Resonance·T F PrisnerS Th Sigurdsson
Mar 13, 2015·Biophysical Journal·Virginia MeyerGareth R Eaton
May 10, 2017·The Journal of Physical Chemistry. B·Indra D SahuGary A Lorigan
Feb 13, 2016·Chemistry : a European Journal·Silvia ValeraBela E Bode
Sep 23, 2014·Biochemistry·Indra D SahuGary A Lorigan
Apr 1, 2016·Physical Chemistry Chemical Physics : PCCP·D AbdullinO Schiemann
Jun 12, 2018·ELife·Erika A RiedererFrancis I Valiyaveetil
Nov 9, 2018·RNA·Nicole ErlenbachThomas F Prisner
May 18, 2020·Biomolecules·Indra D Sahu, Gary A Lorigan
Feb 8, 2019·The Journal of General Physiology·Vanessa LeoneLucy R Forrest
Nov 28, 2012·Physical Chemistry Chemical Physics : PCCP·Andriy Marko, Thomas F Prisner
Apr 3, 2018·BioMed Research International·Indra D Sahu, Gary A Lorigan
Nov 30, 2019·Nucleic Acids Research·Marcel HeinzGerhard Hummer
Dec 24, 2013·The Journal of Biological Chemistry·Benesh JosephEnrica Bordignon
Mar 1, 2021·European Biophysics Journal : EBJ·Igor TkachMarina Bennati
Jan 22, 2021·Accounts of Chemical Research·Austin Gamble JarviSunil Saxena
Apr 20, 2010·The Journal of Physical Chemistry. B·Zhongyu YangSunil Saxena

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