Strategy for the use of affinity grids to prepare non-his-tagged macromolecular complexes for single-particle electron microscopy

Journal of Molecular Biology

Volumn 400, Issue 4, 2010, Pages 675-681

  Kelly, Deborah F ^a   Dukovski, Danijela ^a   Walz, Thomas ^a  

^a HARVARD MEDICAL SCHOOL   (United States)

Author keywords

Affinity Grid; Monolayer purification; Ribosome; RNA polymerase II; Single particle electron microscopy

Indexed keywords

ANTIBODY; CELL EXTRACT; HISTIDINE; PROTEIN A; RNA POLYMERASE II;

AFFINITY GRID; ARTICLE; ELECTRON MICROSCOPY; HUMAN; LIPID MONOLAYER; MACROMOLECULE; PRIORITY JOURNAL; RIBOSOME; SINGLE PARTICLE ELECTRON MICROSCOPY;

EID: 77954384708     PISSN: 00222836     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.jmb.2010.05.045     Document Type: Article

References (23)

1. Frank J. Single-particle reconstruction of biological macromolecules in electron microscopy-30 years. Q. Rev. Biophys. 2009, 42:139-158.
2. Stagg S.M., Lander G.C., Pulokas J., Fellmann D., Cheng A., Quispe J.D., et al. Automated cryoEM data acquisition and analysis of 284742 particles of GroEL. J. Struct. Biol. 2006, 155:470-481.
3. Ludtke S.J., Baldwin P.R., Chiu W. EMAN: semiautomated software for high-resolution single-particle reconstructions. J. Struct. Biol. 1999, 128:82-97.
4. Cheng Y., Walz T. The advent of near-atomic resolution in single-particle electron microscopy. Annu. Rev. Biochem. 2009, 78:723-742.
5. Zhou Z.H. Towards atomic resolution structural determination by single-particle cryo-electron microscopy. Curr. Opin. Struct. Biol. 2008, 18:218-228.
6. Kelly D.F., Dukovski D., Walz T. Monolayer purification: a rapid method for isolating protein complexes for single-particle electron microscopy. Proc. Natl Acad. Sci. USA 2008, 105:4703-4708.
7. Kelly D.F., Abeyrathne P.D., Dukovski D., Walz T. The Affinity Grid: a pre-fabricated EM grid for monolayer purification. J. Mol. Biol. 2008, 382:423-433.
8. Forsgren A., Sjoquist J. "Protein A" from S. aureus. I. Pseudo-immune reaction with human gamma-globulin. J. Immunol. 1966, 97:822-827.
9. Li Z., Hite R.K., Cheng Y., Walz T. Evaluation of imaging plates as recording medium for images of negatively stained single particles and electron diffraction patterns of two-dimensional crystals. J. Electron Microsc. 2009, 59:53-63.
10. Dukovski D., Li Z., Kelly D.F., Mack E., Walz T. Structural and functional studies on the stalk of the transferrin receptor. Biochem. Biophys. Res. Commun. 2009, 381:712-716.
11. Chandramouli P., Topf M., Menetret J.F., Eswar N., Cannone J.J., Gutell R.R., et al. Structure of the mammalian 80S ribosome at 8.7 Å resolution. Structure 2008, 16:535-548.
12. Frank J., Radermacher M., Penczek P., Zhu J., Li Y., Ladjadj M., Leith A. SPIDER and WEB: processing and visualization of images in 3D electron microscopy and related fields. J. Struct. Biol. 1996, 116:190-199.
13. Ohi M., Li Y., Cheng Y., Walz T. Negative staining and image classification-powerful tools in modern electron microscopy. Biol. Proced. Online 2004, 6:23-34.
14. Cheng Y., Wolf E., Larvie M., Zak O., Aisen P., Grigorieff N., et al. Single particle reconstructions of the transferrin-transferrin receptor complex obtained with different specimen preparation techniques. J. Mol. Biol. 2006, 355:1048-1065.
15. Cramer P., Bushnell D.A., Fu J., Gnatt A.L., Maier-Davis B., Thompson N.E., et al. Architecture of RNA polymerase II and implications for the transcription mechanism. Science 2000, 288:640-649.
16. Cramer P., Bushnell D.A., Kornberg R.D. Structural basis of transcription: RNA polymerase II at 2.8 Å resolution. Science 2001, 292:1863-1876.
17. Chung W.H., Craighead J.L., Chang W.H., Ezeokonkwo C., Bareket-Samish A., Kornberg R.D., Asturias F.J. RNA polymerase II/TFIIF structure and conserved organization of the initiation complex. Mol. Cell 2003, 12:1003-1013.
18. Kostek S.A., Grob P., De Carlo S., Lipscomb J.S., Garczarek F., Nogales E. Molecular architecture and conformational flexibility of human RNA polymerase II. Structure 2006, 14:1691-1700.
19. Craighead J.L., Chang W.H., Asturias F.J. Structure of yeast RNA polymerase II in solution: implications for enzyme regulation and interaction with promoter DNA. Structure 2002, 10:1117-1125.
20. Kettenberger H., Armache K.J., Cramer P. Complete RNA polymerase II elongation complex structure and its interactions with NTP and TFIIS. Mol. Cell 2004, 16:955-965.
21. Grigorieff N. FREALIGN: high-resolution refinement of single particle structures. J. Struct. Biol. 2007, 157:117-125.
22. Bottcher B., Wynne S.A., Crowther R.A. Determination of the fold of the core protein of hepatitis B virus by electron cryomicroscopy. Nature 1997, 386:88-91.
23. Gnatt A.L., Cramer P., Fu J., Bushnell D.A., Kornberg R.D. Structural basis of transcription: an RNA polymerase II elongation complex at 3.3 Å resolution. Science 2001, 292:1876-1882.