2013-10-02 OBS 2013-05-28 PDBe PDBe 2013-07-03 2013-10-02 2013-10-02

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Ford RC, Holzenburg A Photosynthesis, thylakoid membrane, CF1-CF0 proton ATPase, photosystem, cytochrome b6f, ribosome, chloroplast. Ford RC, Holzenburg A Organization of protein complexes and a mechanism for grana formation in photosynthetic membranes as revealed by cryo-electron microscopy Crystal Research and Technology n/a n/a n/a 2013 doi:10.1002/crat.201300116

emd_2388.tif

1 Thylakoid membrane from spinach. cellular-component Thylakoid membrane Photosynthetic membrane

Buffer - 20mM MES, 5mM MgCl2, 15mM NaCl, pH6.5, 150mM sorbitol. 3microlitres of membranes at 2mg per ml chlorophyll were directly applied to electron microscope grids (Quantifoil carbon grids with 1.2 micron holes). After blotting excess liquid, (2x 1s blots at 95 percent relative humidity in a Vitrobot device), the grids were rapidly frozen by plunging into liquid ethane and transferred at less than 110K to the Gatan cryo-stage of the electron microscope.

false Spinacia oleracea Spinach Leaf Chloroplast 2 x 1 sec blots before plunging. NITROGEN

Vitrification instrument: Vitrobot FEI. Rapid plunging into liquid ethane.

95 FEI VITROBOT MARK I 97 Astigmatism corrected by live FFT. FIELD EMISSION GUN 45 BRIGHT FIELD 8000 96 8000 FLOOD BEAM 98 Single tilt 2.0 -65 33998 49 97 FEI TECNAI F20 28-JUL-2009 GATAN LIQUID NITROGEN 200 34000 77 15 No stain. tissue 2 Quantifoil 400mesh carbon grids with 1.2 micron holes.

20mM MES, 5mM MgCl2, 15mM NaCl, pH6.5, 150mM sorbitol.

6.5 tomography eTOMO

Standard eTOMO procedures for single axis tilt tomograms. Fiducials were selected from the tomogram and tracked in each image by hand initially.

45 Resolution of known features of complexes C1 1 57

1 degree increments above 45 deg, 2.5 degree increments below this.