1O76

CYANIDE COMPLEX OF P450CAM FROM PSEUDOMONAS PUTIDA

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X12C
Synchrotron site	NSLS
Beamline	X12C
Temperature [K]	100
Detector technology	CCD
Detector	BRANDEIS
Spacegroup name	P 1 21 1
Unit cell lengths	67.000, 61.800, 94.600
Unit cell angles	90.00, 90.40, 90.00

Refinement procedure

Resolution	20.000 * - 1.800
R-factor	0.1964
Rwork	0.196
R-free	0.23600 *
Structure solution method	FOURIER SYNTHESIS
RMSD bond length	0.011
RMSD bond angle	1.930 *
Data reduction software	XDS
Data scaling software	XSCALE
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.000
High resolution limit [Å]	1.800	1.800
Rmerge	0.082 *	0.242 *
Total number of observations	237007 *
Number of reflections	67892
<I/σ(I)>	8.2	4
Completeness [%]	94.5	88.9
Redundancy	3.49	2.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7.6 *	4 *	CRYSTALS WERE GROWN USING THE SITTING DROP GEOMETRY AT 2 DEG. C. 5 UL OF 30 MG/ML P450 IN 50 MM TRIS HCL, 250 MM KCL, 0.5 MM CAMPHOR WERE MIXED WITH AN EQUAL VOLUME OF THE RESERVOIR SOLUTION (27-30% PEG 4000, 100 MM DTE, SAME BUFFER AS PROTEIN)., pH 7.40

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	15 (mg/ml)
10	1	reservoir	dithiothreitol	100 (mM)
11	1	reservoir	Na+-MES	100 (mM)	pH6.5
12	1	reservoir	Na+-malonate	0.7 (M)
2	1	drop	EPPS	40 (mM)	pH7.6
3	1	drop		150 (mM)
4	1	drop	glycerol	5 (%)
5	1	drop	H4B	4000 (nM)
6	1	drop	beta-mercaptoethanol	2 (mM)
7	1	drop	beta-octylglucoside	50 (mM)
8	1	reservoir	Na+-L-Arg	100 (mM)	pH7.0
9	1	reservoir		60 (mM)