2REQ

METHYLMALONYL-COA MUTASE, NON-PRODUCTIVE COA COMPLEX, IN OPEN CONFORMATION REPRESENTING SUBSTRATE-FREE STATE

Experimental procedure

Source type	SYNCHROTRON
Source details	ELETTRA BEAMLINE 5.2R
Synchrotron site	ELETTRA
Beamline	5.2R
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1995-07
Detector	MAR scanner 180 mm plate
Spacegroup name	P 1 21 1
Unit cell lengths	104.950, 162.060, 104.200
Unit cell angles	90.00, 108.75, 90.00

Refinement procedure

Resolution	20.000 - 2.500
Rwork	0.259
R-free	0.32500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1req
RMSD bond length	0.012
RMSD bond angle	0.041
Data reduction software	MOSFLM
Data scaling software	CCP4 ((SCALA))
Phasing software	AMoRE
Refinement software	REFMAC

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.640
High resolution limit [Å]	2.500	2.500
Rmerge	0.092	0.338
Number of reflections	113512
<I/σ(I)>	6	2.1
Completeness [%]	99.8	99.7
Redundancy	4.8	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	7.5	23 *	PROTEIN SOLUTION: 20 MG/ML PROTEIN, 1MM ADENOSYLCOBALAMIN, 2MM COA, 1MM DTT, TRIS -HCL PH 7.5. RESERVOIR SOLUTION: 17.5% (W/V) PEG4000, 20% GLYCEROL (V/V), 100MM TRIS-HCL PH 7.5. EQUAL VOLUMES OF PROTEIN SOLUTION WERE MIXED AND EQUILIBRATED BY VAPOR DIFFUSION., vapor diffusion

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	20 (mg/ml)
2	1	drop	coenzyme B12	1 (mM)
3	1	drop	CoA	2 (mM)
4	1	drop	dithiothreitol	1 (mM)
5	1	drop	Tris-HCl
6	1	reservoir	PEG4000	17.5 (%(w/v))
7	1	reservoir	glycerol	20 (%(v/v))
8	1	reservoir	Tris-HCl	100 (mM)