1W16

rat synaptotagmin 4 C2B domain in the absence of calcium

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-BM
Synchrotron site	APS
Beamline	19-BM
Temperature [K]	100
Detector technology	CCD
Collection date	2003-11-29
Detector	CUSTOM
Spacegroup name	P 63 2 2
Unit cell lengths	91.061, 91.061, 122.860
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	30.000 - 2.300
R-factor	0.217
Rwork	0.213
R-free	0.25700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1k5w
RMSD bond length	0.020
RMSD bond angle	1.851
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	AMoRE
Refinement software	REFMAC (5.2.0003)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	29.810	2.340
High resolution limit [Å]	2.300	2.300
Rmerge	0.053	0.725
Number of reflections	13974
<I/σ(I)>	45.8	1.8
Completeness [%]	99.6	95.2
Redundancy	11.4	4.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5		THE RAT SYNAPTOTAGMIN IV C2B DOMAIN WAS CRYSTALLIZED BY THE VAPOR DIFFUSION METHOD (HANGING DROP) BY MIXING 1 MICROLITER PROTEIN (IN 20 MM MES, 150 MM NACL AND 1 MM EDTA, PH 6.32) WITH 1 MICROLITER RESERVOIR SOLUTION (2.5M NACL, 0.1M CACL2, 0.1M HEPES, PH 7.5) SUSPENDED OVER 500 MICROLITERS RESERVOIR SOLUTION. HEXAGONAL CRYSTALS APPEARED OVERNIGHT AND GREW TO A FINAL SIZE OF 0.1 X 0.1 X 0.2 MM WITHIN TWO DAYS. PRIOR TO DATA COLLECTION, CRYSTALS WERE TRANSFERRED INTO CRYSTALLIZATION SOLUTION CONTAINING 4.25 M NACL, 0.1 M HEPES PH 7.5 AND 5%(V/V) ETHYLENE GLYCOL AND THEN COOLED IN LIQUID PROPANE.