Retrospect and Prospect of Single Particle Cryo-Electron Microscopy: The Class of Integral Membrane Proteins as an Example

2020 ◽  
Vol 60 (5) ◽  
pp. 2448-2457 ◽  
Author(s):  
Shirin Akbar ◽  
Sukanya Mozumder ◽  
Jayati Sengupta
Keyword(s):  
Electron Microscopy ◽  
Membrane Proteins ◽  
Single Particle ◽  
Integral Membrane Proteins ◽  
Cryo Electron Microscopy

scholarly journals Single-particle cryo-EM—How did it get here and where will it go

Science ◽  
2018 ◽  
Vol 361 (6405) ◽  
pp. 876-880 ◽  
Author(s):  
Yifan Cheng
Keyword(s):  
Electron Microscopy ◽  
Structural Biology ◽  
Single Particle ◽  
Electron Crystallography ◽  
The Past ◽  
Cryo Electron Microscopy ◽  
Technological Advances ◽  
The Future ◽  
Electron Cryotomography

Cryo–electron microscopy, or simply cryo-EM, refers mainly to three very different yet closely related techniques: electron crystallography, single-particle cryo-EM, and electron cryotomography. In the past few years, single-particle cryo-EM in particular has triggered a revolution in structural biology and has become a newly dominant discipline. This Review examines the fascinating story of its start and evolution over the past 40-plus years, delves into how and why the recent technological advances have been so groundbreaking, and briefly considers where the technique may be headed in the future.

scholarly journals In situ structure determination using single particle cryo-electron microscopy images

2020 ◽  
Author(s):  
Jing Cheng ◽  
Bufan Li ◽  
Long Si ◽  
Xinzheng Zhang
Keyword(s):  
Electron Microscopy ◽  
Structure Determination ◽  
Single Particle ◽  
Structure Refinement ◽  
Target Function ◽  
Target Protein ◽  
Particle Analysis ◽  
Cryo Electron Microscopy ◽  
Tilt Series

AbstractCryo-electron microscopy (cryo-EM) tomography is a powerful tool for in situ structure determination. However, this method requires the acquisition of tilt series, and its time consuming throughput of acquiring tilt series severely slows determination of in situ structures. By treating the electron densities of non-target protein as non-Gaussian distributed noise, we developed a new target function that greatly improves the efficiency of the recognition of the target protein in a single cryo-EM image without acquiring tilt series. Moreover, we developed a sorting function that effectively eliminates the false positive detection, which not only improves the resolution during the subsequent structure refinement procedure but also allows using homolog proteins as models to recognize the target protein. Together, we developed an in situ single particle analysis (isSPA) method. Our isSPA method was successfully applied to solve structures of glycoproteins on the surface of a non-icosahedral virus and Rubisco inside the carboxysome. The cryo-EM data from both samples were collected within 24 hours, thus allowing fast and simple structural determination in situ.

scholarly journals Cryo-electron Microscopy of Membrane Proteins

2013 ◽  
pp. 325-341 ◽  
Author(s):  
Kenneth N. Goldie ◽  
Priyanka Abeyrathne ◽  
Fabian Kebbel ◽  
Mohamed Chami ◽  
Philippe Ringler ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Membrane Proteins ◽  
Cryo Electron Microscopy

scholarly journals A general mechanism of ribosome dimerization revealed by single-particle cryo-electron microscopy

2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Linda E. Franken ◽  
Gert T. Oostergetel ◽  
Tjaard Pijning ◽  
Pranav Puri ◽  
Valentina Arkhipova ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Single Particle ◽  
General Mechanism ◽  
Cryo Electron Microscopy

How Good Can Single-Particle Cryo-EM Become? What Remains Before It Approaches Its Physical Limits?

2019 ◽  
Vol 48 (1) ◽  
pp. 45-61 ◽  
Author(s):  
Robert M. Glaeser
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Single Particle ◽  
Energy Use ◽  
Optimal Choice ◽  
Structural Basis ◽  
Detective Quantum Efficiency ◽  
Quantum Metrology ◽  
Cryo Electron Microscopy ◽  
Induced Motion

Impressive though the achievements of single-particle cryo–electron microscopy are today, a substantial gap still remains between what is currently accomplished and what is theoretically possible. As is reviewed here, twofold or more improvements are possible as regards ( a) the detective quantum efficiency of cameras at high resolution, ( b) converting phase modulations to intensity modulations in the image, and ( c) recovering the full amount of high-resolution signal in the presence of beam-induced motion of the specimen. In addition, potential for improvement is reviewed for other topics such as optimal choice of electron energy, use of aberration correctors, and quantum metrology. With the help of such improvements, it does not seem to be too much to imagine that determining the structural basis for every aspect of catalytic control, signaling, and regulation, in any type of cell of interest, could easily be accelerated fivefold or more.

scholarly journals Structure of the Blm10-20S Proteasome Complex by Single Particle Cryo Electron Microscopy

2006 ◽  
Vol 12 (S02) ◽  
pp. 372-373
Author(s):  
J Ortega ◽  
J Iwanczyk ◽  
K Sadre-Bazzaz ◽  
K Ferrell ◽  
E Kondrashkina ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Single Particle ◽  
20S Proteasome ◽  
Cryo Electron Microscopy

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

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