2014-11-26 REL 2014-10-15 PDBe PDBe 2014-10-29 2014-10-29 single-particle cryo reconstruction of the Large Ribosomal subunit-associated protein Quality Control (RQC) complex in the context of Tae2 deletion Lyumkis D, Oliveira dos Passos D, Tahara EB, Webb K, Bennett EJ, Vinterbo S, Potter CS, Carragher B, Joazeiro CAP RING domain E3 ubiquitin ligase, translational surveillance, protein quality control, cryo-EM, listerin/Ltn1, Tae2/Nemf Lyumkis D, Oliveira dos Passos D, Tahara EB, Webb K, Bennett EJ, Vinterbo S, Potter CS, Carragher B, Joazeiro CAP Structural basis for translational surveillance by the large ribosomal subunit-associated protein quality control complex PROC.NAT.ACAD.SCI.USA 111 15981 15986 2014 25349383 doi:10.1073/pnas.1413882111 emd_2798.map.gz Image stored as Reals 128 128 128 0 0 0 127 127 127 128 128 128 432.64 432.64 432.64 90.0 90.0 90.0 X Y Z -2.46616793 9.38885403 0.02034449 0.99826878 1
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3.38 3.38 3.38 1.5 reconstruction of RQC complex in the context of Tae2 protein deletion 1 Large 60S ribosomal subunit in complex with protein quality control components, in the context of Tae2 protein deletion 2.5 ribosome-eukaryote Large 60S ribosomal subunit 2.5 LSU 60S false Saccharomyces cerevisiae Baker's yeast BY4741 protein Large ribosomal subunit-associated quality control complexlex RQC false BY4741 Saccharomyces cerevisiae Baker's yeast specimens were prepared for cryo-EM by applying 3 microliters of sample to a freshly plasma-cleaned holey carbon C-flat grid (Protochips) that had been overlaid with 2 nm thin carbon film, allowing the sample to adsorb to the grid for 30 s, followed by blotting with filter paper and plunge freezing into liquid ethane using a manual cryoplunger in an ambient environment at 4 C. ETHANE HOMEMADE PLUNGER 93 objective lens astigmatism was corrected by monitoring power spectra of continuously acquired images in leginon FIELD EMISSION GUN 32 BRIGHT FIELD 500 4000 FLOOD BEAM 626 TVIPS TEMCAM-F416 (4k x 4k) 2.0 92307 93 FEI TECNAI F20 22-MAY-2013 GATAN LIQUID NITROGEN 200 62000 2048 15.6 particle 0.1 freshly plasma-cleaned holey carbon C-flat grid (Protochips) that had been overlaid with 2 nm thin carbon film
50 mM Hepes-KOH, 100 mM KOAc, 5 mM MgOAc, 1 mM EDTA, 2 mM DTT, 2x protease inhibitors, 0.1% Igepal CA-630
6.8 singleParticle projection-matching Frealign each particle 11.2 FSC 0.143, gold-standard C1 37819
see detailed methods in paper