reSeArCH Letter
a) b)
e) f)
c)
Marker
kDa^13 18 19 2017161514 21 22 23 2425 26
8032
25
17
11(^00102030)
200
400
600
Volume (ml)
A^280
(mAU)
Superdex S200 10/300 GL
Ve=13.1ml
5 10 15 18-20 25 30
153A
Crystal structure of SidJ/CaM fitted into the Cryo EM map
SidJ
Yeast CaM
d)
Resolution (1/Å).
FSC gold standard
Phase randomized
Fourier shell correlation (FSC
)
.
FSC=0.143
n=3154 n=2445
n=2027
n=6565
n=2949
n=2821
n=4065
n=2709
n=6512
n=8607
n=2934
n=4159 n=4117
n=6554
n=5358
n=2241
Extended Data Fig. 6 | Cryo-EM data processing and 3D reconstruction.
a, Size-exclusion profile of SidJ–CaM complex. Elution fractions were
analysed by SDS–PAGE. Marked fractions were used for cryo-EM sample
preparation. This experiment was repeated three times independently,
with similar results. b, A representative electron micrograph for the
cryo-EM dataset collected. c, Reference-free representative 2D class
averages of the SidJ–CaM complex. Secondary structure features are
visible in projection images. The number (n) of particles used to obtain a
2D class average is mentioned in each subpanel. d, Gold-standard FSC^30
plot between two independently refined half-maps, FSC0.143 = 4.15-Å
resolution. As expected, FSC between phase-randomized half-maps
show a rapid drop of correlation beyond randomization point. e, Crystal
structure of SidJ–CaM (PDB 6OQQ) is fitted into the cryo-EM 3D
reconstruction. f, Part of d, magnified to highlight the difference between
the crystal structure and the cryo-EM map.