1BU6

CRYSTAL STRUCTURES OF ESCHERICHIA COLI GLYCEROL KINASE AND THE MUTANT A65T IN AN INACTIVE TETRAMER: CONFORMATIONAL CHANGES AND IMPLICATIONS FOR ALLOSTERIC REGULATION

X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, HANGING DROP	8.5		20-22% (W/V) POLYETHYLENE GLYCOL MR 4000 0.2 M LISO4 0.1 M TRIS PH 8.5 - 8.8 CRYSTALLIZED BY HANGING DROP VAPOR DIFFUSION AT ROOM TEMPERATURE., vapor diffusion - hanging drop

Crystal Properties
Matthews coefficient	Solvent content
2.65	53.1

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 91.9	α = 90
b = 119	β = 103.4
c = 109.3	γ = 90

Symmetry
Space Group	P 1 21 1

Diffraction

Diffraction Experiment
ID #	Crystal ID	Scattering Type	Data Collection Temperature	Detector	Detector Type	Details	Collection Date	Monochromator	Protocol
1	1	x-ray	298	AREA DETECTOR	SDMS		1994-08-15	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	ROTATING ANODE	RIGAKU RU200

Data Collection

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (Observed)	R Merge I (Observed)	Net I Over Average Sigma (I)	Redundancy	Number Reflections (All)	Number Reflections (Observed)	Observed Criterion Sigma (F)	Observed Criterion Sigma (I)	B (Isotropic) From Wilson Plot
1	2.37	20	74	0.068		2		69682		2	32.3

Refinement

Statistics
Diffraction ID	Structure Solution Method	Starting model	Resolution (High)	Resolution (Low)	Number Reflections (Observed)	Number Reflections (R-Free)	Percent Reflections (Observed)	R-Work	R-Free	Mean Isotropic B
X-RAY DIFFRACTION	MOLECULAR REPLACEMENT	1GLF	2.37	20	69682		74	0.167

Temperature Factor Modeling
Anisotropic B[1][1]	Anisotropic B[1][2]	Anisotropic B[1][3]	Anisotropic B[2][2]	Anisotropic B[2][3]	Anisotropic B[3][3]

RMS Deviations
Key	Refinement Restraint Deviation
t_dihedral_angle_d	20.528
t_it	9.34
t_angle_deg	2.928
t_nbd	0.032
t_bond_d	0.019
t_gen_planes	0.019
t_trig_c_planes	0.018
t_incorr_chiral_ct
t_pseud_angle

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	15621
Nucleic Acid Atoms
Solvent Atoms	140
Heterogen Atoms	54

Software

Software
Software Name	Purpose
FRFSUM	model building
UNPUBLISHED	model building
TNT	refinement
SDMS	data reduction
SDMS	data scaling
FRFSUM	phasing
UNPUBLISHED	phasing

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM133198.