1FMO

CRYSTAL STRUCTURE OF A POLYHISTIDINE-TAGGED RECOMBINANT CATALYTIC SUBUNIT OF CAMP-DEPENDENT PROTEIN KINASE COMPLEXED WITH THE PEPTIDE INHIBITOR PKI(5-24) AND ADENOSINE

Experimental procedure

Source type	ROTATING ANODE
Source details	RIGAKU RUH2R
Temperature [K]	277
Detector technology	AREA DETECTOR
Collection date	1994-02-07
Detector	XUONG-HAMLIN MULTIWIRE
Spacegroup name	P 21 21 21
Unit cell lengths	73.080, 78.440, 80.490
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	10.000 - 2.200
R-factor	0.182 *
Rwork	0.182
Structure solution method	ISOMORPHOUS REPLACEMENT
Starting model (for MR)	1apm
RMSD bond length	0.012
RMSD bond angle	2.000 *
Data reduction software	UCSD
Data scaling software	UCSD
Phasing software	X-PLOR (3.1)
Refinement software	TNT (5E)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.300
High resolution limit [Å]	2.200	2.200
Rmerge	0.064 *	0.250 *
Number of reflections	21679
<I/σ(I)>	7.3	1
Completeness [%]	92.0	84
Redundancy	2.4	1.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	4 *	ORTHORHOMBIC CRYSTALS OF THE TERNARY COMPLEX, COMPRISED OF MOUSE HIS6-RC SUBUNIT, PKI (5-24) AND ADENOSINE, WERE GROWN BY THE HANGING-DROP VAPOUR DIFFUSION METHOD USING 2-METHYL-2,4-PENTANEDIOL (MPD) AS THE PRECIPITATING AGENT. THE MOTHER-LIQUOR CONTAINED PROTEIN AT A CONCENTRATION OF 0.25 MM IN 100 MM BICINE BUFFER AT PH 8.0, 0.75 MM PKI(5-24), 4 MM ADENOSINE AND 4% MPD. THE RESERVOIR SOLUTION WAS MADE UP OF 20% MPD IN 100 MM BICINE BUFFER (PH 8.0). X-RAY DIFFRACTION QUALITY CRYSTALS (0.2 X 0.15 X 0.8 MM3) WERE GROWN AT 4 C IN APPROXIMATELY 4 - 6 WEEKS., vapor diffusion - hanging drop, temperature 277K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	0.25 (mM)
2	1	drop	Bicine	100 (mM)
3	1	drop	PKI(5-24)	0.75 (mM)
4	1	drop	adenosine	4 (mM)
5	1	drop	MPD	4 (%)
6	1	reservoir	MPD	20 (%)
7	1	reservoir	Bicine	100 (mM)