2018-11-28 REL PDBe PDBe PDBe 2018-06-13 2018-07-04 2018-08-01 2018-11-28 Masked Core Centromere Binding Factor 3 (CBF3) with monomeric Ndc10 Zhang WJ Lukoynova N Zhang W Lukoynova N Miah S Lucas J Vaughan CK Insights into Centromere DNA Bending Revealed by the Cryo-EM Structure of the Core Centromere Binding Factor 3 with Ndc10. Cell Rep US 24 744 754 2018 30021170 doi:10.1016/j.celrep.2018.06.068 2211-1247 EMD-4241 other EM volume
Core CBF3 complex without Ndc10
 EMD-0051 other EM volume
0051 is generated from the same data with refinement of the complete map
 EMD-0052 associated EM volume Core CBF3 in complex with Ndc10 D1-2 Core CBF3 in complex with Ndc10 D1-2 0 1 2 3 4
The truncated CBF3 complex, recombinantly expressed in Saccharomyces cerevisiae. It comprises a Cep3 homodimer, in which the binuclear zinc cluster domains are truncated, full length heterodimer of Skp1 and Ctf13, and a monomeric construct Ndc10 comprising domains 1-2.
0.286 Cep3 homodimer 1 1 Saccharomyces cerevisiae Saccharomyces cerevisiae Skp1 1 2 Saccharomyces cerevisiae Saccharomyces cerevisiae modified pRS424 and pRS426 Ctf13 1 3 Saccharomyces cerevisiae Saccharomyces cerevisiae Ndc10 comprising domains 1-2 1 4 Saccharomyces cerevisiae Saccharomyces cerevisiae singleParticle particle 0.15 8. 200.0 NaCl sodium chloride 15.0 Tris base 2.0 DTT ETHANE 100 277 FEI VITROBOT MARK I
The sample is homogeneous and well-dispersed on grids.
FEI TITAN KRIOS FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 300 2.7 47170. FEI TITAN KRIOS AUTOGRID HOLDER NITROGEN GATAN K2 SUMMIT (4k x 4k) COUNTING 1-40 1 2003 50.0 1
The selected images were high-pass filtered and normalized.
214608
Number of particles after selection by 2D classification
 CTFFIND 4.0 CTF parameters were estimated using CTFFIND4
Cryosparc was used to generate a startup model from a smaller dataset collected in-house. This was then low pass filtered to 60 Angstroms
 2 C1 FOURIER SPACE 4.4 FSC 0.143 CUT-OFF RELION 2.0
This map was generated using focused refinement of the Ndc10 component of the map.
22965 ANGULAR RECONSTITUTION RELION 2.0 ANGULAR RECONSTITUTION RELION 2.0 6 RELION 2.0 emd_0052.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 256 256 256 0 0 0 256 256 256 271.36 271.36 271.36 90.0 90.0 90.0 X Y Z -0.034055203 0.092103794 0.0005317477 0.0039967815 1.06 1.06 1.06 0.0221 AUTHOR focused refinement of Ndc10D1-2 in CBF3
"Fit in map" function used to place 6FE8 and 4ACO with out further refinement or model building