A Refined Single-Particle Reconstruction Procedure to Process Two-Dimensional Crystal Images from Transmission Electron Microscopy

2015 ◽  
Vol 21 (4) ◽  
pp. 876-885 ◽  
Author(s):  
Qie Kuang ◽  
Pasi Purhonen ◽  
Thirupathi Pattipaka ◽  
Yohannes H. Ayele ◽  
Hans Hebert ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Membrane Proteins ◽  
Single Particle ◽  
Three Dimensional ◽  
Reconstruction Procedure ◽  
Single Particle Reconstruction ◽  
Secondary Transporter ◽  
Unit Cells ◽  
2D Crystals ◽  
Particle Reconstruction

AbstractSingle-particle reconstruction (SPR) and electron crystallography (EC), two major applications in electron microscopy, can be used to determine the structure of membrane proteins. The three-dimensional (3D) map is obtained from separated particles in conventional SPR, but from periodic unit cells in EC. Here, we report a refined SPR procedure for processing 2D crystal images. The method is applied to 2D crystals of melibiose permease, a secondary transporter inEscherichia coli. The current procedure is improved from our previously published one in several aspects. The “gold standard Fourier shell correlation” resolution of our final reconstruction reaches 13 Å, which is significantly better than the previously obtained 17 Å resolution. The choices of different refinement parameters for reconstruction are discussed. Our refined SPR procedure could be applied to determine the structure of other membrane proteins in small or locally distorted 2D crystals, which are not ideal for EC.

scholarly journals Development of Nanostructured Electron Microscopy Grids for Time Resolved Single Particle Reconstruction for Transmission Electron Microscopy

2006 ◽  
Vol 12 (S02) ◽  
pp. 1130-1131
Author(s):  
D Barnard ◽  
TR Shaikh ◽  
K Buttle ◽  
V Pushparaj ◽  
D Ye ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Single Particle ◽  
Time Resolved ◽  
Single Particle Reconstruction ◽  
Transmission Electron ◽  
Particle Reconstruction

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2005

Single particle reconstruction of membrane proteins: A tool for understanding the 3D structure of disease-related macromolecules

2010 ◽  
Vol 103 (1) ◽  
pp. 122-130 ◽  
Author(s):  
Kazuhiro Mio ◽  
Yuusuke Maruyama ◽  
Toshihiko Ogura ◽  
Masaaki Kawata ◽  
Toshio Moriya ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Single Particle ◽  
3D Structure ◽  
Single Particle Reconstruction ◽  
Particle Reconstruction

scholarly journals Single-particle reconstruction of biological macromolecules in electron microscopy – 30 years

2009 ◽  
Vol 42 (3) ◽  
pp. 139-158 ◽  
Author(s):  
Joachim Frank
Keyword(s):  
Electron Microscopy ◽  
Single Particle ◽  
Specimen Preparation ◽  
Biological Macromolecules ◽  
Personal Account ◽  
Dynamic Information ◽  
Preparation Methods ◽  
Single Particle Reconstruction ◽  
Macromolecular Assemblies ◽  
Particle Reconstruction

AbstractThis essay gives the autho's personal account on the development of concepts underlying single-particle reconstruction, a technique in electron microscopy of macromolecular assemblies with a remarkable record of achievements as of late. The ribosome proved to be an ideal testing ground for the development of specimen preparation methods, cryo-EM techniques, and algorithms, with discoveries along the way as a rich reward. Increasingly, cryo-EM and single-particle reconstruction, in combination with classification techniques, is revealing dynamic information on functional molecular machines uninhibited by molecular contacts.

scholarly journals Imaging Three-Dimensional Elemental Inhomogeneity in Pt–Ni Nanoparticles Using Spectroscopic Single Particle Reconstruction

Nano Letters ◽  
2019 ◽  
Vol 19 (2) ◽  
pp. 732-738 ◽  
Author(s):  
Yi-Chi Wang ◽  
Thomas J. A. Slater ◽  
Gerard M. Leteba ◽  
Alan M. Roseman ◽  
Christopher P. Race ◽  
...  
Keyword(s):  
Single Particle ◽  
Three Dimensional ◽  
Ni Nanoparticles ◽  
Single Particle Reconstruction ◽  
Particle Reconstruction

Single Particle Reconstruction of Yeast RNA Degradation Machine, Exosome, Using Electron Microscopy

2008 ◽  
Vol 14 (S2) ◽  
pp. 162-163
Author(s):  
HW Wang ◽  
J Wang ◽  
F Ding ◽  
K Callahan ◽  
MA Bratkowski ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
New Mexico ◽  
Single Particle ◽  
Rna Degradation ◽  
Single Particle Reconstruction ◽  
Particle Reconstruction

Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008

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