Sedimentation Yields Long-Term Stable Protein Samples as Shown by Solid-State NMR

Frontiers in Molecular Biosciences
Thomas WiegandAnja Böckmann

Abstract

Today, the sedimentation of proteins into a magic-angle spinning (MAS) rotor gives access to fast and reliable sample preparation for solid-state Nuclear Magnetic Resonance (NMR), and this has allowed for the investigation of a variety of non-crystalline protein samples. High protein concentrations on the order of 400 mg/mL can be achieved, meaning that around 50-60% of the NMR rotor content is protein; the rest is a buffer solution, which includes counter ions to compensate for the charge of the protein. We have demonstrated herein the long-term stability of four sedimented proteins and complexes thereof with nucleotides, comprising a bacterial DnaB helicase, an ABC transporter, an archaeal primase, and an RNA polymerase subunit. Solid-state NMR spectra recorded directly after sample filling and up to 5 years later indicated no spectral differences and no loss in signal intensity, allowing us to conclude that protein sediments in the rotor can be stable over many years. We have illustrated, using an example of an ABC transporter, that not only the structure is maintained, but that the protein is still functional after long-term storage in the sedimented state.

References

Nov 1, 1989·Pharmaceutical Research·M C ManningR T Borchardt
Jan 1, 1993·Annual Review of Physical Chemistry·J L Silva, G Weber
Mar 1, 1996·Protein Engineering·E QuerolA Mozo-Villarias
Jun 20, 1997·Journal of Molecular Biology·G VogtP Argos
Dec 28, 1999·Proceedings of the National Academy of Sciences of the United States of America·N HillsonA E García
Jan 5, 2000·Journal of Magnetic Resonance·B M FungK Ermolaev
May 29, 2002·Nature Structural Biology·Rasmus FoghErnest Laue
Mar 15, 2006·Annals of the New York Academy of Sciences·John C Bischof, Xiaoming He
May 27, 2006·Proceedings of the National Academy of Sciences of the United States of America·Matthew W BowlerJohn E Walker
Sep 1, 2006·Nature·Roger J P Dawson, Kaspar P Locher
Oct 9, 2007·Journal of Magnetic Resonance·Peter L Gor'kovWilliam W Brey
Sep 26, 2009·Journal of Biomolecular NMR·Anja BöckmannAnne Lesage
Feb 10, 2010·Pharmaceutical Research·Mark Cornell ManningDerrick S Katayama
Jun 15, 2011·Proceedings of the National Academy of Sciences of the United States of America·Ivano BertiniPaola Turano
Sep 29, 2011·Journal of Biomolecular NMR·Tim J StevensErnest D Laue
Nov 29, 2011·Physical Chemistry Chemical Physics : PCCP·Ivano BertiniPaola Turano
Apr 13, 2012·Proceedings of the National Academy of Sciences of the United States of America·Julien RocheCatherine A Royer
Sep 8, 2012·Journal of the American Chemical Society·Yaqiang WangGary J Pielak
Mar 9, 2013·Accounts of Chemical Research·Ivano BertiniEnrico Ravera
Aug 31, 2013·Journal of Biomolecular NMR·Marco FragaiEnrico Ravera
Sep 7, 2013·Critical Reviews in Food Science and Nutrition·Wangang ZhangDong U Ahn
May 13, 2015·Proceedings of the National Academy of Sciences of the United States of America·Guilherme A P de Oliveira, Jerson L Silva
May 20, 2015·Computational and Structural Biotechnology Journal·Julien BoudetGeorg Lipps
Aug 19, 2015·Biomolecular NMR Assignments·Thomas WiegandBeat H Meier
Dec 15, 2015·Frontiers in Veterinary Science·Pietro Celi, Gianfranco Gabai
Jan 13, 2016·Angewandte Chemie·Enrico RaveraClaudio Luchinat
Mar 31, 2016·The Biochemical Journal·Michael J Davies
Oct 25, 2016·Biomolecular NMR Assignments·Dennis W PiehlChad M Rienstra
Feb 24, 2017·Journal of Biomolecular NMR·Abhishek MandalPatrick C A van der Wel
Dec 5, 2017·Journal of Biomolecular NMR·Denis LacabanneAnja Böckmann
Feb 22, 2018·Angewandte Chemie·ShengQi XiangHugo van Ingen
Mar 16, 2018·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Lauriane LecoqAnja Böckmann
Apr 19, 2018·Proceedings of the National Academy of Sciences of the United States of America·Cyril CharlierAdriaan Bax
Jun 19, 2018·Emerging Topics in Life Sciences·Patrick C A van der Wel
Oct 6, 2018·Journal of the American Chemical Society·Hundeep KaurClemens Glaubitz
Feb 26, 2019·Progress in Nuclear Magnetic Resonance Spectroscopy·Denis LacabanneAnja Böckmann

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Citations

Jun 24, 2020·Biochemical Society Transactions·Arun Kumar Sreemantula, Alexander Marchanka
Nov 18, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Denis LacabanneAnja Böckmann
Sep 30, 2020·Viruses·Lauriane LecoqAnja Böckmann
Apr 22, 2021·Proceedings of the National Academy of Sciences of the United States of America·Lauriane LecoqAnja Böckmann
Apr 7, 2021·Chemistry : a European Journal·Johannes ZehnderThomas Wiegand
Jun 8, 2021·The Journal of Physical Chemistry. B·Alexander A MalärThomas Wiegand
Nov 27, 2020·The Journal of Physical Chemistry. B·Denis LacabanneThomas Wiegand
Sep 5, 2021·Progress in Nuclear Magnetic Resonance Spectroscopy·Giacomo ParigiClaudio Luchinat
Sep 8, 2021·Nature Communications·Alexander A MalärThomas Wiegand
Sep 30, 2021·Annual Review of Virology·Gal Porat-DahlerbruchTatyana Polenova

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Methods Mentioned

BETA
Nuclear Magnetic Resonance
NMR
X-ray
deamination
gel-filtration

Software Mentioned

CcpNmr
TOPSPIN

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