2BIF

6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE-2,6-BISPHOSPHATASE H256A MUTANT WITH F6P IN PHOSPHATASE ACTIVE SITE

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	120
Detector technology	IMAGE PLATE
Collection date	1995-03-15
Detector	RIGAKU
Spacegroup name	P 1
Unit cell lengths	61.740, 73.510, 76.700
Unit cell angles	116.90, 99.31, 105.20

Refinement procedure

Resolution	30.000 - 2.400
R-factor	0.2
Rwork	0.200
R-free	0.24400
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bif
RMSD bond length	0.012
RMSD bond angle	23.800 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	X-PLOR
Refinement software	X-PLOR (3.851)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	2.050
High resolution limit [Å]	2.000	2.000
Rmerge	0.052 *
Number of reflections	37144 *
<I/σ(I)>	21.6	3.7
Completeness [%]	95.1 *	60.6
Redundancy	2.16 *	1.56

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7.5 *	4 *	90MM SUCCINATE, PH 6.0, 17% PEG4000, 1% B-OCTYLGLUCOSIDE, 10% GLYCEROL, pH 7.0

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	10 (mg/ml)
10	1	drop	EDTA	0.040 (mM)
11	1	reservoir	succinate	60-90 (mM)
12	1	reservoir	glycerol	10 (%)
13	1	reservoir		10 (mM)
14	1	reservoir	PEG4000	17 (%)
2	1	drop	Tris-PO4	50 (mM)
3	1	drop	glycerol	5 (%)
4	1	drop	EGTA	0.040 (mM)
5	1	drop	dithiothreitol	0.800 (mM)
6	1	drop	PEG300	0.4 (%)
7	1	drop	Fru-6-P	1.7 (mM)
8	1	drop	AMP-PNP	1.7 (mM)
9	1	drop	beta-octylglucoside	1 (%)