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Self-Perforated Hydrogel Nanomembranes Facilitate Structural Analysis Of Proteins By Electron Cryo-Microscopy.

Julian Scherr, K. Parey, Niklas Klusch, B. J. Murphy, Sebastian Balser, Alexander Neuhaus, V. Zickermann, W. Kühlbrandt, A. Terfort, Daniel Rhinow
Published 2017 · Materials Science, Medicine

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We developed a method to improve specimen preparation for electron cryo-microscopy of membrane proteins. The method features a perforated hydrogel nanomembrane that stabilizes the thin film of aqueous buffer spanning the holes of holey carbon films, while at the same time preventing the depletion of protein molecules from these holes. The membrane is obtained by cross-linking of thiolated polyglycerol dendrimer films on gold, which self-perforate upon transfer to holey carbon substrates, forming a sub-micron-sized hydrogel network. The perforated nanomembrane improves the distribution of the protein molecules in the ice considerably. This facilitates data acquisition as demonstrated with two eukaryotic membrane protein complexes.
This paper references
10.1016/j.jsb.2012.02.003
Beam-induced motion of vitrified specimen on holey carbon film.
Axel F. Brilot (2012)
10.1002/jcc.20084
UCSF Chimera—A visualization system for exploratory research and analysis
E. F. Pettersen (2004)
10.1038/nmeth.2472
Electron counting and beam-induced motion correction enable near atomic resolution single particle cryoEM
Xueming Li (2013)
10.1039/b923756a
Ultrathin conductive carbon nanomembranes as support films for structural analysis of biological specimens.
Daniel Rhinow (2010)
10.1016/j.jsb.2012.09.006
RELION: Implementation of a Bayesian approach to cryo-EM structure determination
S. Scheres (2012)
10.1016/j.str.2012.08.026
Movies of ice-embedded particles enhance resolution in electron cryo-microscopy.
M. G. Campbell (2012)
10.1006/JSBI.1999.4174
EMAN: semiautomated software for high-resolution single-particle reconstructions.
S. Ludtke (1999)
10.1021/bm801093t
Synthesis and characterization of glycerol dendrons, self-assembled monolayers on gold: a detailed study of their protein resistance.
M. Wyszogrodzka (2009)
10.1038/nmeth.2931
Controlling protein adsorption on graphene for cryo-EM using low-energy hydrogen plasmas
C. J. Russo (2014)
10.1016/j.jsb.2009.12.020
Graphene oxide: a substrate for optimizing preparations of frozen-hydrated samples.
R. Pantelic (2010)
10.1002/CHEM.200306073
Self-assembled monolayers of dendritic polyglycerol derivatives on gold that resist the adsorption of proteins.
Conrad Siegers (2004)
New Insights into the Unique Structure of the F 0 F 1-ATP Synthase from the Chlamydomonad Algae Polytomella sp . and Chlamydomonas reinhardtii 1
R. Lis (2007)
10.1016/j.colsurfb.2013.05.026
Direct grafting of anti-fouling polyglycerol layers to steel and other technically relevant materials.
T. Weber (2013)
10.1021/CM010756C
Hydrophilic Oligo(oxyethylene)-Derivatized Poly(3,4-ethylenedioxythiophenes): Cation-Responsive Optoelectroelectrochemical Properties and Solid-State Chromism
I. F. Perepichka (2002)
10.1038/nmeth.2115
Prevention of overfitting in cryo-EM structure determination
S. Scheres (2012)
10.1016/j.jsb.2013.01.004
A method to achieve homogeneous dispersion of large transmembrane complexes within the holes of carbon films for electron cryomicroscopy
M. Cheung (2013)
10.1038/nature14185
Horizontal membrane-intrinsic α-helices in the stator a-subunit of an F-type ATP synthase
M. Allegretti (2015)
10.1016/j.ultramic.2009.04.002
Detective quantum efficiency of electron area detectors in electron microscopy
G. McMullan (2009)
10.1146/annurev-biochem-070511-103700
Mitochondrial complex I.
J. Hirst (2013)
10.1038/srep07084
Self-assembled monolayers improve protein distribution on holey carbon cryo-EM supports
Joel R. Meyerson (2014)
10.1016/S1047-8477(03)00069-8
Accurate determination of local defocus and specimen tilt in electron microscopy.
J. A. Mindell (2003)
10.1016/S0005-2728(00)00266-8
Efficient large scale purification of his-tagged proton translocating NADH:ubiquinone oxidoreductase (complex I) from the strictly aerobic yeast Yarrowia lipolytica.
N. Kashani-Poor (2001)
10.1016/J.JSB.2005.03.010
Automated molecular microscopy: the new Leginon system.
C. Suloway (2005)
10.1016/j.jsb.2011.09.002
Initial evaluation of a direct detection device detector for single particle cryo-electron microscopy.
Anna-Clare Milazzo (2011)
10.1016/j.bbabio.2016.09.014
Cryo-EM structure of respiratory complex I reveals a link to mitochondrial sulfur metabolism.
Edoardo D'Imprima (2016)
10.1021/MA0340762
Hyperbranched Polyglycidol on Si/SiO2 Surfaces via Surface-Initiated Polymerization
Majad Khan (2003)
10.1016/j.ultramic.2007.11.005
Electron cryo-microscopy of biological specimens on conductive titanium-silicon metal glass films.
D. Rhinow (2008)
Page 19 of 22 ACS Paragon Plus Environment ACS Nano 1 2 3 4 5
Rasband (1997)
10.1126/science.1259530
Ultrastable gold substrates for electron cryomicroscopy
C. J. Russo (2014)
10.1016/j.jsb.2010.10.002
Graphene: Substrate preparation and introduction.
R. Pantelic (2011)
10.1038/nmeth.4193
MotionCor2: anisotropic correction of beam-induced motion for improved cryo-electron microscopy
Shawn Zheng (2017)
10.1038/nmeth.2727
The Local Resolution of Cryo-EM Density Maps
A. Kucukelbir (2014)
Preventing Overfitting in Cryo-EM Structure Determination Nat
S. H. Scheres (2012)
10.1016/j.jsb.2015.08.008
CTFFIND4: Fast and accurate defocus estimation from electron micrographs.
A. Rohou (2015)
10.1126/science.1251652
The Resolution Revolution
W. Kühlbrandt (2014)
10.1021/la702867t
Self-assembled monothiol-terminated hyperbranched polyglycerols on a gold surface: a comparative study on the structure, morphology, and protein adsorption characteristics with linear poly(ethylene glycol)s.
Po-Ying J Yeh (2008)
10.1016/j.ultramic.2013.06.004
High-resolution noise substitution to measure overfitting and validate resolution in 3D structure determination by single particle electron cryomicroscopy☆
S. Chen (2013)
10.1109/ISBI.2012.6235807
A Bayesian view on cryo-EM structure determination
S. Scheres (2012)



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