1P16

Structure of an mRNA capping enzyme bound to the phosphorylated carboxyl-terminal domain of RNA polymerase II

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X9A
Synchrotron site	NSLS
Beamline	X9A
Temperature [K]	100
Detector technology	CCD
Collection date	2001-10-15
Detector	MARRESEARCH
Wavelength(s)	0.9792
Spacegroup name	P 1 21 1
Unit cell lengths	47.112, 124.970, 92.705
Unit cell angles	90.00, 100.54, 90.00

Refinement procedure

Resolution	20.000 * - 2.700
R-factor	0.2001
Rwork	0.200
R-free	0.26600
Structure solution method	SAD
RMSD bond length	0.008
RMSD bond angle	1.099 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	SOLVE
Refinement software	CNS (0.9)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.800
High resolution limit [Å]	2.700	2.700
Rmerge	0.074	0.427
Total number of observations	495195 *
Number of reflections	56988 *
<I/σ(I)>	10.8	2.8
Completeness [%]	96.2	89.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion *	8 *	291	3M ammonium phosphate, 50mM hepes, 3% DMSO, pH 7.3, VAPOR DIFFUSION, HANGING DROP, temperature 291K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	10 (mg/ml)
2	1	drop		60 (mM)
3	1	drop	Tris-HCl	20 (mM)	pH8.0
4	1	drop	beta-mercaptoethanol	1 (mM)
5	1	reservoir	ammonium phosphate	3.0 (M)
6	1	reservoir	HEPES	50 (mM)	pH7.3
7	1	reservoir	DMSO	3 (%(v/v))