mitotic checkpoint complex / meiotic sister chromatid cohesion, centromeric / Inactivation of APC/C via direct inhibition of the APC/C complex / APC/C:Cdc20 mediated degradation of mitotic proteins / anaphase-promoting complex / metaphase/anaphase transition of mitotic cell cycle / outer kinetochore / protein localization to chromosome, centromeric region / mitotic spindle assembly checkpoint signaling / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal ...mitotic checkpoint complex / meiotic sister chromatid cohesion, centromeric / Inactivation of APC/C via direct inhibition of the APC/C complex / APC/C:Cdc20 mediated degradation of mitotic proteins / anaphase-promoting complex / metaphase/anaphase transition of mitotic cell cycle / outer kinetochore / protein localization to chromosome, centromeric region / mitotic spindle assembly checkpoint signaling / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / Resolution of Sister Chromatid Cohesion / APC-Cdc20 mediated degradation of Nek2A / RHO GTPases Activate Formins / Cdc20:Phospho-APC/C mediated degradation of Cyclin A / kinetochore / spindle / Separation of Sister Chromatids / non-specific serine/threonine protein kinase / protein kinase activity / cell division / protein serine kinase activity / protein serine/threonine kinase activity / centrosome / apoptotic process / perinuclear region of cytoplasm / ATP binding / nucleus / cytosol / cytoplasm Similarity search - Function
Serine Threonine Protein Phosphatase 5, Tetratricopeptide repeat - #430 / Mad3/Bub1 homology region 1 / Mitotic spindle checkpoint protein Bub1/Mad3 / Mad3/BUB1 homology region 1 / BUB1 N-terminal domain profile. / Mad3/BUB1 hoMad3/BUB1 homology region 1 / Serine Threonine Protein Phosphatase 5, Tetratricopeptide repeat / Alpha Horseshoe / Protein kinase-like domain superfamily / Mainly Alpha Similarity search - Domain/homology
ACETATE ION / PRASEODYMIUM ION / Mitotic checkpoint serine/threonine-protein kinase BUB1 beta Similarity search - Component
Biological species
HOMO SAPIENS (human)
Method
X-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 1.8 Å
Mass: 18.015 Da / Num. of mol.: 117 / Source method: isolated from a natural source / Formula: H2O
Nonpolymer details
ETHANE-1,2-DIOL (EDO): USED AS CRYOPROTECTANT PRASEODYMIUM ION (PR): ESSENTIAL FOR HIGH RESOLUTION ...ETHANE-1,2-DIOL (EDO): USED AS CRYOPROTECTANT PRASEODYMIUM ION (PR): ESSENTIAL FOR HIGH RESOLUTION OF CRYSTALS AND USED FOR PHASING
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Experimental details
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Experiment
Experiment
Method: X-RAY DIFFRACTION / Number of used crystals: 2
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Sample preparation
Crystal
Density Matthews: 2.47 Å3/Da / Density % sol: 49 % / Description: THIS DATA SET WAS USED IN REFINEMENTS.
Crystal grow
pH: 8 Details: 22.5% (W/V) PEG 8000, 0.17 M TRIS PH 8.0, 0.01 M PRASEODYMIUM(III) ACETATE
Type: ADSC CCD / Detector: CCD / Date: Oct 12, 2007 Details: KIRKPATRICK BAEZ BIMORPH MIRROR PAIR FOR HORIZONTAL AND VERTICAL FOCUSSING
Radiation
Monochromator: SI (111) DOUBLE CRYSTAL MONOCHROMATOR / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelength
Wavelength: 1.06 Å / Relative weight: 1
Reflection
Resolution: 1.8→46.62 Å / Num. obs: 19685 / % possible obs: 99.9 % / Observed criterion σ(I): 2 / Redundancy: 15.6 % / Biso Wilson estimate: 22 Å2 / Rmerge(I) obs: 0.11 / Net I/σ(I): 21
Reflection shell
Resolution: 1.8→1.9 Å / Redundancy: 15.8 % / Rmerge(I) obs: 0.51 / Mean I/σ(I) obs: 4 / % possible all: 100
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Processing
Software
Name
Version
Classification
REFMAC
5.2.0019
refinement
MOSFLM
datareduction
SCALA
datascaling
autoSHARP
phasing
Refinement
Method to determine structure: SAD Starting model: NONE Resolution: 1.8→54.39 Å / Cor.coef. Fo:Fc: 0.943 / Cor.coef. Fo:Fc free: 0.93 / SU B: 3.444 / SU ML: 0.106 / Cross valid method: THROUGHOUT / ESU R: 0.139 / ESU R Free: 0.135 / Stereochemistry target values: MAXIMUM LIKELIHOOD Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. DENSITY WAS NOT OBSERVED FOR RESIDUES 92-95. ONE ROUND OF TLS WAS CONDUCTED DURING REFINEMENT. TWO GROUPS WERE USED; RESIDUES 57-114 AND 115-220.
Rfactor
Num. reflection
% reflection
Selection details
Rfree
0.25986
1005
5.1 %
RANDOM
Rwork
0.22188
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obs
0.22373
18643
99.91 %
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Solvent computation
Ion probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK