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Title: Cryo-EM structure of the 60S-Rei1 complex Authors: Greber BJ, Boehringer D, Montellese C, Ban N Sample: 60S ribosomal subunit in complex with Rei1 Method: Single particle reconstruction (14.6 angstroms resolution) Latest update: 2012-12-12   Summary  Experimental details   Visualization   Map information   Downloads Sample Sample name: 60S ribosomal subunit in complex with Rei1 Oligomeric state: One monomer each of 60S and Rei1 Theoretical molecular weight of the sample: 2.4 Components: ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details 1 ribosome-eukaryote 60S ribosomal subunit false Saccharomyces cerevisiae 2 protein Rei1 monomer true Saccharomyces cerevisiae IPR007087, IPR015880 C-terminal His-tag Experiment Specimen state: Particle Specimen preparation: pH Specimen conc. Details Staining Specimen support details 8.3 mg/mL 20 mM Hepes-NaOH pH 8.3, 50 mM NaCl, 5 mM MgCl2, 5 mM beta-mercaptoethanol cryo Quantifoil holey carbon grid R2/1 Vitrification: Cryogen name Humidity Temp. Instr. Method Time resolved Details ETHANE % 80 K HOMEMADE PLUNGER manual blotting ms Vitrification instrument: manual plunger Imaging: Microscope Voltage Illumination mode Imaging mode Cs Defocus min. Defocus max. Nominal mag. Calibrated mag. Electron source Detector Detector distance Astigmatism FEI TECNAI 20 200 kV FLOOD BEAM BRIGHT FIELD 2.3 mm 1300 nm 6000 nm 83000 FIELD EMISSION GUN Gatan Ultrascan 4000 mm Specimen holder Holder model Tilt min. Tilt max. Energy filter Energy window Temp. Temp. min. Temp. max. Beam tilt Electron dose Other details Date eucentric GATAN LIQUID NITROGEN 0° 0° eV 87 K K K mrad 20 e/Å2 08-FEB-2012 Processing Protocol: projection matching Software: SPIDER CTF correction: per frame Number of particles: 17415 Resolution by author: 14.6 Å Resolution method: FSC at 0.5 cut-off Processing details: Fourier amplitudes of the reconstruction were enhanced using the SAXS curve from ribosomes; subsequently, the map was filtered in SPIDER using a butterworth low-pass filter with a filter midpoint of 0.22. Scanned images: Num. images Sampling size OD range Quant. bit number Other details Scanner 15 μm/pixel 16 images were acquired using a 2 x 2 frame spot scan (per hole) using a serial EM script Fitting: PDB Protocol Target crit. Software B value Fitting space PDB chain Details 3U5H  rigid body correlation Chimera REAL Protocol: rigid body 3U5I  rigid body correlation Chimera REAL Protocol: rigid body