REL PDBe PDBe PDBe 2004-05-13 2004-05-25 2005-05-25 2012-10-17 The PM2 virion has a novel organization with an internal membrane and pentameric receptor binding spikes. Huiskonen JT Kivela HM Bamford DH Butcher SJ Huiskonen JT Kivela HM Bamford DH Butcher SJ The PM2 virion has a novel organization with an internal membrane and pentameric receptor binding spikes. NAT.STRUCT.MOL.BIOL. 11 850 856 2004 15286721 doi:10.1038/nsmb807 Proteinase K-treated PM2 virus Proteinase K-treated PM2 virus
Purified virion (1 mg/ml) was treated with 75 micrograms/ml proteinase K, purified by sucrose rate zonal ulatracentrifugation, pelleted and resuspended just prior to application to the grid.
1 32.6 Reported mass of the whole virion in the literature is 47 MDa, 72 percent of which is protein. Mass spectrometry was used to find the protease cleavage site on the spike protein. Estimated copy number was 60 for this protein. A mass of 1.2 MDa was thus deducted from the theoretical MW of the virion Pseudoalteromonas phage PM2
includes DNA, lipid and proteins. Protease K-treated
Pseudoalteromonas phage PM2 Pseudoalteromonas sp. ER72M2 BACTERIA(EUBACTERIA) 45.8 P2 597 21 VIRION STRAIN true false protease K-treated PM2 singleParticle particle 4 7.2
CaCl2, 5mM Tris-HCl, 10-20 mM NaCl, 25-125 mM
400 mesh copper grid, R2/2 quantifoil holey
 ETHANE 90 HOMEMADE PLUNGER
Vitrification instrument: EMBL design
A small vial of ethane is placed inside a larger liquid nitrogen reservoir. The grid holding 3 microliters of the sample is held in place at the bottom of a plunger by the means of fine tweezers. When the liquid ethane is ready, a piece of filter paper is then pressed against the sample to blot off excess buffer, sufficient to leave a thin layer on the grid. The filter paper is removed, and the plunger is allowed to drop into the liquid ethane. Once the grid enters the liquid ethane, the sample is rapidly frozen, and the grid is transferred under liquid nitrogen to a storage box immersed in liquid nitrogen for later use in the microscope. FEI TECNAI F20 FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 200 2 1.1 2.2 50000.0 GATAN LIQUID NITROGEN 90 94 93 objective lens astigmatism was corrected at
A box anticontaminator was fitted with a minimum achievable temperature of 88 K
KODAK SO-163 FILM ZEISS SCAI 7 8 10 12. Side entry liquid nitrogen-cooled cryo specimen holder
The particles were first automatically selected using ETHAN, and then boxed out manually in EMAN.
each particle, wiener factor 0.1
 I OTHER 13.0 FSC 0.5 CUT-OFF EMBL, P3DR 752 emd_1084.map.gz 1 IMAGE STORED AS SIGNED INTEGER (2 BYTES) 325 325 325 0 0 0 325 325 325 910 910 910 90 90 90 X Y Z -2930.0 4431.0 0.185507 680.033999999999992 2.8 2.8 2.8 1110.0 This is the reconstruction of the PM2 virion after treatment with protease K has cleaved the spike protein
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