HPUB OBS PDBe PDBe PDBe 2013-06-25 2013-07-24 2014-07-09 2014-07-09 Negative stain electron microscopy reconstruction of the tip complex from the type III secretion system of Shigella flexneri with MxiH mutation Q51A Cheung M Shen D Makino F Kato T Fernando VK Roehrich D Martinez-Argudo I Walker ML Liu X Brown J Frazer G Mantell J Mina P Todd T Sessions RB Namba K Blocker AJ Tip complex, Type III secretion system, Shigella flexneri, Q51A Cheung M Shen D Makino F Kato T Fernando VK Roehrich D Martinez-Argudo I Walker ML Liu X Brown J Frazer G Mantell J Mina P Todd T Sessions RB Namba K Blocker AJ Structure and activation mechanism of the type III secretion system needle tip complex To Be Published Veenendaal AK Hodgkinson JL Schwarzer L Stabat D Zenk SF Blocker AJ The type III secretion system needle tip complex mediates host cell sensing and translocon insertion MOL.MICROBIOL. 63 1719 1730 2007 17367391 doi:10.1111/j.1365-2958.2007.05620.x Blocker AJ Deane JE Veenendaal AK Roversi P Hodgkinson JL Johnson S Lea SM What's the point of the type III secretion system needle? PROC.NAT.ACAD.SCI.USA 105 6507 6513 2008 18458349 doi:10.1073/pnas.0708344105 Conformation of the tip complex from the type III secretion system of Shigella flexneri bound to the needle with MxiH mutation Q51A Conformation of the tip complex from the type III secretion system of Shigella flexneri bound to the needle with MxiH mutation Q51A
Tip complexes were analysed from purified needle complexes and therefore when still bound to the needle tip
Five subunits of the tip complex bound to fifty subunits of the needle 2 0.68 Tip complex 1 Heteropentameric GO:0030257 IPR006135 true Shigella flexneri serotype 5a Type III secretion system Extracellular component of membrane embedded complex 0.21 Shigella flexneri M90T derivative pACT3, pBAD Needle 1 GO:0030257 IPR006135 true Shigella flexneri serotype 5a Type III secretion system Extracellular component of membrane embedded complex 0.47 Shigella flexneri M90T derivative pACT3, pBAD singleParticle particle 0.1 8.0
0.1% (w/v) DDM, 150 mM NaCl, 25 mM Tris pH 8, 5 mM EDTA
 NEGATIVE
Carbon coating mica sheets were submerged in a drop of 0.01% (w/v) DDM until the carbon sheet partially detached from the mica. The mica was then submerged in the sample for 5 s, then submerged in a drop of water for 5 s. The mica was then placed onto the surface of a drop of 1% uranyl acetate and allowed to sink, causing the carbon to float on the surface of the drop. After 10 s, the carbon sheet was adsorbed onto a grid.
300 mesh copper grid with a thin carbon film to which the specimen was adsorbed to
 NONE OTHER FEI TECNAI 20 FLOOD BEAM BRIGHT FIELD LAB6 200 2 1.4 1.4 50000.0 70754.0 SIDE ENTRY, EUCENTRIC Thon rings visualised at 50-100,000 times magnification using the fast FT and corrected accordingly 0 2011-08-31 FEI EAGLE (4k x 4k) 15 250 9 460 3.56 16.0 FEI single tilt holder
Particles were selected using helix boxer from EMAN2. After alignment, Euler angles were determined by projection matching with projections of a previously determined 16A resolution map of the needle. The reconstruction was iterated and helical pattern searching and imposition was applied to each new reference generated after each iteration.
First zero set to 18 angstrom, no CTF correction not required
 OTHER 22.5 OTHER EMAN2, SPIDER, bespoke-2D-alignment, bespoke-X-and-Y-shifting 2545
SPIDER: theta 4 degrees
 127 emd_2406.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 200 200 200 0 0 0 200 200 200 423.99997 423.99997 423.99997 90.0 90.0 90.0 X Y Z -0.07658133 0.10239315 0.00017214 0.00833076 2.12 2.12 2.12 0.0332 AUTHOR Negative stain electron microscopy reconstruction of the tip complex and needle from the type III secretion system of Shigella flexneri with MxiH mutation Q51A
::::EMDATABANK.org::::EMD-2406::::
 2J0N A RIGID BODY FIT Chimera
Residues comprising the N domain were removed. The 10 residues at the C terminus (not resolved in the crystal structure) were built into an alpha helix
REAL emd_2406_fsc.xml