2014-09-10 REL 2008-09-29 PDBe PDBe 2008-09-29 2010-06-01 Recognition of Phe-tRNAPhe, Trp-tRNATrp and : a common molecular mechanism for aminoacyl-tRNA selection revealed by cryo-EM Li W, Agirrezabala X, Lei J, Bouakaz L, Brunelle JL, Ortiz-Meoz RF, Green R, Sanyal S, Ehrenberg M, Frank J protein translation, decoding, elongation factor Tu, ribosome 3eq4 Li W, Agirrezabala X, Lei J, Bouakaz L, Brunelle JL, Ortiz-Meoz RF, Green R, Sanyal S, Ehrenberg M, Frank J Recognition of aminoacyl-tRNA: a common molecular mechanism revealed by cryo-EM. EMBO J. 27 3322 3331 2008 19020518 doi:10.1038/emboj.2008.243 emd_1564.map.gz Image stored as Reals 130 130 130 -65 -65 -65 64 64 64 130 130 130 366.6 366.6 366.6 90 90 90 X Y Z -0.000495627 0.00147413 3.53844e-05 0.000183878 1
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2.82 2.82 2.82 0.00045 Leu-tRNA-EFTu-kir-GDP-70S ribosome 5 Leu-tRNA-EFTu-GDP-kir-70S ribosome monomeric 2.8
sedimentation
2.8 ribosome-prokaryote 70S ribosome 70S ribosome 2.8 2.8 ALL Escherichia coli false Blot for 6 seconds NITROGEN
Vitrification instrument: vitrobot
90 FEI VITROBOT 80 corrected objective at 100,000 times magnification FIELD EMISSION GUN 15 no energy filter BRIGHT FIELD 1000 80.7 4000 FLOOD BEAM 80.7 cartridge KODAK SO-163 FILM 2.26 0 49645 0 80.7 FEI TECNAI F20 01-JUN-2006 no tilt OTHER 300 50000 ZEISS SCAI 112 7 2
Protocol: real space refinement. 1lBM(1989-1996 fragment)
TNT rigid body REAL 2AVY 2AW4 1GIX 1IBM 1QZA cryo 0.03 200 mesh particle
Polymix buffer( 5 mM potassium phosphate (KH2PO4) pH 7.3, 5 mM NH4Cl, 95 mM KCl, 0.5 mM CaCl2, 8 mM putrescine, 1 mM spermidine, 1 mM DTE, 5 mM magnesium acetate)
7.3 singleParticle weighted back-projection spider Spider defocus groups, Wiener filter 12 FSC 0.5 80000 C1