2OA1

Crystal Structure of RebH, a FAD-dependent halogenase from Lechevalieria aerocolonigenes, the L-Tryptophan with FAD complex

X-RAY DIFFRACTION

Starting Model(s)

Initial Refinement Model(s)
Type	Source	Accession Code	Details
experimental model	PDB	2O9Z	apo form of same protein in same lattice

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, HANGING DROP		277	Protein Solution (18 mg/ml protein, 0.050 M sodium chloride, 0.010 M TRIS pH 8.0) mixed in a 1:1 ratio with the Well Solution (0.9 M K2HPO4, 0.5 M NaH2PO4) crystals soaked for 22 hours in solution of 0.6 M K2HPO4, 0.33 M NaH2PO4, ~0.005 M FAD, ~0.003 M L-tryptophan, 0.030 M NaCl, Cryoprotected with: well solution supplemented with up to 30% glycerol, VAPOR DIFFUSION, HANGING DROP, temperature 277K

Crystal Properties
Matthews coefficient	Solvent content
3.5	64.7

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 114.492	α = 90
b = 114.492	β = 90
c = 231.935	γ = 120

Symmetry
Space Group	P 62

Diffraction

Diffraction Experiment
ID #	Crystal ID	Scattering Type	Data Collection Temperature	Detector	Detector Type	Details	Collection Date	Monochromator	Protocol
1	1	x-ray	100	CCD	MARMOSAIC 300 mm CCD	Adjustable focusing mirrors in K-B geometry	2006-06-18	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	APS BEAMLINE 23-ID-D	0.97931	APS	23-ID-D

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	2.15	49.576	98.8	0.102	24.905	20.4		92078

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	R Merge I (Observed)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
1	2.15	2.2	90.9		0.534	3.628	9.4	5634

Refinement

Statistics
Diffraction ID	Structure Solution Method	Cross Validation method	Starting model	Resolution (High)	Resolution (Low)	Number Reflections (Observed)	Number Reflections (R-Free)	Percent Reflections (Observed)	R-Factor (Observed)	R-Work	R-Free	R-Free Selection Details	Mean Isotropic B
X-RAY DIFFRACTION	2O9Z	THROUGHOUT	apo form of same protein in same lattice	2.15	19.952	91914	4630	98.846	0.154	0.152	0.194	RANDOM	38.03

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]
0.031	0.015		0.031		-0.046

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	32.395
r_dihedral_angle_4_deg	15.397
r_dihedral_angle_3_deg	13.511
r_dihedral_angle_1_deg	6.171
r_scangle_it	3.39
r_scbond_it	2.086
r_angle_refined_deg	1.412
r_mcangle_it	1.157
r_mcbond_it	0.725
r_nbtor_refined	0.304

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	32.395
r_dihedral_angle_4_deg	15.397
r_dihedral_angle_3_deg	13.511
r_dihedral_angle_1_deg	6.171
r_scangle_it	3.39
r_scbond_it	2.086
r_angle_refined_deg	1.412
r_mcangle_it	1.157
r_mcbond_it	0.725
r_nbtor_refined	0.304
r_symmetry_hbond_refined	0.245
r_nbd_refined	0.194
r_symmetry_vdw_refined	0.184
r_xyhbond_nbd_refined	0.149
r_chiral_restr	0.098
r_bond_refined_d	0.015
r_gen_planes_refined	0.006

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	8468
Nucleic Acid Atoms
Solvent Atoms	1056
Heterogen Atoms	103

Software

Software
Software Name	Purpose
DENZO	data reduction
SCALEPACK	data scaling
REFMAC	refinement
PDB_EXTRACT	data extraction
HKL-2000	data reduction

RCSB PDB Core Operations are funded by the U.S. National Science Foundation (DBI-2321666), 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 R01GM157729.