Ligand Induced Conformational Changes of a Membrane Transporter in E. coli Cells Observed with DEER/PELDOR

Journal of the American Chemical Society
Benesh JosephDavid S Cafiso

Abstract

An unrealized goal in structural biology is the determination of structure and conformational change at high resolution for membrane proteins within the cellular environment. Pulsed electron-electron double resonance (PELDOR) is a well-established technique to follow conformational changes in purified membrane protein complexes. Here we demonstrate the first proof of concept for the use of PELDOR to observe conformational changes in a membrane protein in intact cells. We exploit the fact that outer membrane proteins usually lack reactive cysteines and that paramagnetic spin labels entering the periplasm are selectively reduced to achieve specific labeling of the cobalamin transporter BtuB in Escherichia coli. We characterize conformational changes in the second extracellular loop of BtuB upon ligand binding and compare the PELDOR data with high-resolution crystal structures. Our approach avoids detergent extraction, purification, and reconstitution usually required for these systems. With this approach, structure, function, conformational changes, and molecular interactions of outer membrane proteins can be studied at high resolution in the cellular environment.

References

Feb 17, 1999·Archives of Biochemistry and Biophysics·N RapoportA A Timoshin
Jan 29, 2000·Journal of Magnetic Resonance·M PannierH W Spiess
Mar 26, 2003·Nature Structural Biology·David P ChimentoMichael C Wiener
Oct 24, 2006·The Journal of Biological Chemistry·Li MaPhillip E Klebba
Dec 23, 2006·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·Ranjan ChakrabortyDick van der Helm
Jun 15, 2007·Quarterly Reviews of Biophysics·Olav Schiemann, Thomas F Prisner
Jun 21, 2007·Molecular Microbiology·Surendranathan Devanathan, Kathleen Postle
Jun 2, 2010·Journal of the American Chemical Society·Ryuji IgarashiMasahiro Shirakawa
Sep 2, 2010·Journal of Magnetic Resonance·Richard WardDavid G Norman
Dec 1, 2010·Physical Chemistry Chemical Physics : PCCP·Yevhen PolyhachGunnar Jeschke
Apr 21, 2011·Angewandte Chemie·Ivan KrstićThomas F Prisner
May 24, 2011·Biochemical Society Transactions·William Dowhan, Mikhail Bogdanov
Dec 6, 2011·Annual Review of Physical Chemistry·Mei HongFanghao Hu
Mar 13, 2012·Annual Review of Physical Chemistry·Gunnar Jeschke
Jun 9, 2012·Journal of the American Chemical Society·Zhongyu YangWayne L Hubbell
Jul 4, 2012·Physical Chemistry Chemical Physics : PCCP·Yevhen PolyhachGunnar Jeschke
Jan 10, 2013·The Journal of Physical Chemistry Letters·Peter P BorbatJack H Freed
Dec 24, 2013·The Journal of Biological Chemistry·Benesh JosephEnrica Bordignon
Jan 17, 2014·Journal of the American Chemical Society·Moritz J SchmidtDaniel Summerer
Apr 12, 2014·Current Opinion in Structural Biology·Jacob Piehler
Jun 26, 2014·Journal of the American Chemical Society·Georgiy Yu ShevelevElena G Bagryanskaya
Jul 30, 2014·Angewandte Chemie·Robert HänselVolker Dötsch
Aug 29, 2014·Journal of the American Chemical Society·Andrea MartoranaDaniella Goldfarb
Oct 18, 2014·Journal of the American Chemical Society·Mian QiMalte Drescher
Mar 13, 2015·Biophysical Journal·Virginia MeyerGareth R Eaton
Nov 6, 2015·The Journal of Physical Chemistry Letters·Philipp E SpindlerThomas F Prisner

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Citations

Nov 3, 2016·Biophysical Journal·Arthur SikoraDavid S Cafiso
Dec 30, 2016·The Journal of Organic Chemistry·Shengdian HuangAndrzej Rajca
Mar 16, 2017·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Matthew J LawlessSunil Saxena
May 18, 2016·Organic & Biomolecular Chemistry·P RoserD Summerer
Jun 28, 2019·Chembiochem : a European Journal of Chemical Biology·Alessio BonucciElisabetta Mileo
May 18, 2020·Biomolecules·Indra D Sahu, Gary A Lorigan
Mar 23, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·J Jacques JassoyOlav Schiemann
Jan 23, 2020·Proceedings of the National Academy of Sciences of the United States of America·Laura GalazzoEnrica Bordignon
Jan 19, 2019·International Journal of Molecular Sciences·Theresa BraunDaniel Summerer
Feb 4, 2021·Physical Chemistry Chemical Physics : PCCP·Joshua L WortBela E Bode
Jun 18, 2017·Archives of Biochemistry and Biophysics·Rivkah Rogawski, Ann E McDermott
May 1, 2019·Journal of Magnetic Resonance·Kelsey M McCoyAnn E McDermott
Mar 16, 2021·The Journal of Physical Chemistry Letters·Joshua L WortBela E Bode
Mar 25, 2021·Biochimica Et Biophysica Acta. Proteins and Proteomics·F TorricellaA Bonucci
Apr 1, 2019·Biochimica Et Biophysica Acta. General Subjects·Tomohide Saio, Koichiro Ishimori
May 18, 2021·The Journal of Physical Chemistry. B·Katrin AckermannBela E Bode
Jan 9, 2019·The Journal of Physical Chemistry. B·Yin YangDaniella Goldfarb
Feb 28, 2020·The Journal of Organic Chemistry·Haraldur Y Juliusson, Snorri Th Sigurdsson
Apr 2, 2020·The Journal of Physical Chemistry. B·Zhimin YangAndrzej Rajca
Nov 19, 2021·Chemical Communications : Chem Comm·Anandi KugeleMalte Drescher

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