Sample name: R107G mutation of small heat shock protein (sHSP) HSP16.5 from Methanocaldococcus jannaschii (MjHSP16.5)
Protein: MjHSP16.5 R107G mutation
Applied symmetry: O
Number of particles: 5010
Number of class averages: 193
Reconstruction protocol: projection matching
CTF correction: Each image
Details: The final reconstructed density map was further sharpened by application of an amplitude correction algorithm in the program BFACTOR The particles were selected using an automatic selection program Gautomatch developed in Fei Sun lab (to be published). Octahedron symmetry were imposed during 3D reconstructing.
Microscope model: FEI TITAN KRIOS
Electron source: FIELD EMISSION GUN
Electron dose (e/A**2): 20.0
Nominal CS (mm): 2.70
Nominal magnification: 96000.0
Defocus max (nm): 3,000.00
Defocus min (nm): 2,000.00
Holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Number of images: 1308
Details: Electron micrograph exposures were made with the automatic collection package Leginon.
Specimen state: particle
Specimen concentration (mg/mL): 0.80
Buffer: 10mM HEPES, 100mM NaCl.
Specimen support: GIG holey grids (LifeTrust, China) were treated with a glow discharge machine (Master Plasmer)
Apparatus: FEI VITROBOT MARK IV
Protocol: The samples were heated to 60 degree Celsius in boiler, then 3.5 microliter samples were added to the grid and blotted for 2 s with blot force 2 at 90% humidity.