2017-02-01 OBS 2015-05-28 PDBe PDBe 2015-07-15 2017-02-01 2017-02-01
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EMD-3029 Single particle electron microscopy of the type 6 secretion system complex TssA C-terminal fragment Zoued A, Durand E, Spinelli S, Brunet YR, Douzi B, Bebeacua C, Legrand P, Guzzo M, Journet L, Mignot T, Cambillau C, Cascales E Single particle, EM, TssA, C-terminal, type 6 secretion system Zoued A, Durand E, Spinelli S, Brunet YR, Douzi B, Bebeacua C, Legrand P, Guzzo M, Journet L, Mignot T, Cambillau C, Cascales E Caught in the act: TssA primes and leads polymerization of the Type VI secretion tail tube and sheath To Be Published emd_3030.map.gz Image stored as Reals 64 64 64 9 9 9 72 72 72 64 64 64 225.92 225.92 225.92 90.0 90.0 90.0 X Y Z -0.06170310 0.22169326 0.00732864 0.02944504 1
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3.53 3.53 3.53 0.05 Reconstruction of the C-terminal fragment of the TssA complex
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1 TssA complex - C-terminal fragment dodecamer 0.181 protein TssA C-terminus 0.181
The complex appears as a dodecamer but presents 6-fold symmetry.
Escherichia coli BL21 true Dodecamer 1 pETG-20A Escherichia coli BL21 NONE NONE LAB6 10 BRIGHT FIELD FLOOD BEAM GENERIC CCD 2.2 60000 FEI TECNAI SPIRIT 01-NOV-2012 SIDE ENTRY, EUCENTRIC 120 60000 50 The sample was incubated on a previously glow-discharged carbon grid at a concentration of 0.05 mgml-1 for 1 minute. Excess solution was blotted away and the grid was then stained with 2% uranyl acetate and left to air dry after the blotting away excess staining. particle 0.05 Carbon coated grids.
Tris-HCl 20 mM, NaCl 150 mM.
8 singleParticle Maximum Likelihood Xmipp, Spider 36 FSC 1/2 BIT, semi-independent 500 C6 5000
Particles were selected using the program boxer from the EMAN2 package, extracted into boxes and combined into a dataset. The dataset was pre-treated using the SPIDER package and submitted to maximum likelihood (ML) classification and alignment using the Xmipp package. The aligned datasets were then classified by principal component analysis (PCA) using SPIDER into classes with approximately 10 members per class. The initial models were built from class averages selected as a side-views imposing 6-fold symmetry. Due to artefacts resulting from the excess numbers of top-view images, the dataset was reduced to their side-images components during refinement. The initial model was then refined by three-dimensional ML refinement over the reduced dataset imposing C6 symmetry.