Visualization of macromolecular complexes using cryo-electron microscopy with FEI Tecnai transmission electron microscopes

Nature Protocols

Volumn 3, Issue 2, 2008, Pages 330-339

  Grassucci, Robert A ^a   Taylor, Derek ^b   Frank, Joachim ^a,c  

^a Columbia University College of Physicians and Surgeons ^*  (United States)

^b WADSWORTH CENTER   (United States)

^c Och Spine at New York Presbyterian Hospitals   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; COMPLEX FORMATION; CONTROLLED STUDY; CRYOELECTRON MICROSCOPY; ELECTRON BEAM; FROZEN SECTION; IMAGE ANALYSIS; IMAGE PROCESSING; IMAGE RECONSTRUCTION; INFORMATION PROCESSING; MACROMOLECULE; PRIORITY JOURNAL; STANDARD; THREE DIMENSIONAL IMAGING; TRANSMISSION ELECTRON MICROSCOPY;

CRYOELECTRON MICROSCOPY; IMAGE PROCESSING, COMPUTER-ASSISTED; IMAGING, THREE-DIMENSIONAL; MACROMOLECULAR SUBSTANCES; MICROSCOPY, ELECTRON, TRANSMISSION;

EID: 39449126462     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2007.474     Document Type: Article

References (14)

1. Grassucci, R.A., Taylor, D.J. & Frank, J. Preparation of macromolecular complexes for cryo-electron microscopy. Nat. Protoc. 2, 3239-3246 (2007).
2. Frank, J. Three-Dimensional Electron Microscopy of Macromolecular Assemblies: Visualization of Biological Molecules in Their Native State. (Oxford University Press, New York, 2006).
3. Dubochet, J., Lepault, J., Freeman, R., Berriman, J.A. & Homo, J.-C. Electron microscopy of frozen water and aqueous solutions. J. Microsc. 128, 219-237 (1982).
4. Wagenknecht, T., Carazo, J.M., Radermacher, M. & Frank, J. Three-dimensional reconstruction of the ribosome from Escherichia coli. Biophys. J. 55, 455-464 (1989).
5. Spahn, C.M. et al. Hepatitis C virus IRES RNA-induced changes in the conformation of the 40S ribosomat subunit. Science 291, 1959-1962 (2001).
6. Valle, M. et al. Incorporation of aminoacyl-tRNA into the ribosome as seen by cryo-electron microscopy. Nat. Struct. Biol. 10, 899-906 (2003).
7. Halic, M. et al. Structure of the signal recognition particle interacting with the elongation-arrested ribosome. Nature 427 808-814 (2004).
8. Radermacher, M., Wagenknecht, T., Verschoor, A. & Frank, J. Three-dimensional reconstruction from a single-exposure, random conical tilt series applied to the 50S ribosomal subunit of Escherichia coli. J. Microsc. 146, 113-136 (1987).
9. Taylor, D.J. et al. Structures of modified eEF2 80S ribosome complexes reveal the role of GTP hydrolysis in translocation. EMBO J. 26, 2421-2431 (2007).
10. Penczek, P.A., Zhu, J. & Frank, J. A common-lines based method for determining orientations for N > 3 particle projections simultaneously. Ultramicroscopy 63, 205-218 (1996).
11. Van Heel, M. Angular reconstitution: a posteriori assignment of projection directions for 3D reconstruction. Ultramicroscopy 21 111-123 (1987).
12. Lei, J. & Frank, J. Automated acquisition of cryo-electron micrographs for single particle reconstruction on an FEI Tecnai electron microscope. J. Struct. Biol. 150, 69-80 (2005).
13. Suloway, C. et al. Automated molecular microscopy: The new Leginon system. J. Struct. Biol. 151, 41-60 (2005).
14. Ohi, M., Li, Y., Cheng, Y. & Walz, T. Negative staining and image classification - powerful tools in modern electron microscopy. Biol. Proced. Online 6, 23-34 (2004).