3ZYM

Structure of CALM (PICALM) in complex with VAMP8

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I03
Synchrotron site	Diamond
Beamline	I03
Temperature [K]	100
Detector technology	CCD
Collection date	2010-10-20
Detector	ADSC CCD
Spacegroup name	C 1 2 1
Unit cell lengths	161.051, 100.260, 104.021
Unit cell angles	90.00, 118.88, 90.00

Refinement procedure

Resolution	45.573 - 2.030
R-factor	0.1815
Rwork	0.181
R-free	0.19800
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1hf8
RMSD bond length	0.008
RMSD bond angle	0.913
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	PHENIX ((PHENIX.REFINE))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	91.100	2.080
High resolution limit [Å]	2.000	2.030
Rmerge	0.080	0.730
Number of reflections	93133
<I/σ(I)>	10	1.9
Completeness [%]	99.7	99.8
Redundancy	3.7	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.2	289	CRYSTALS WERE GROWN IN SITTING DROPS CONTAINING EQUAL AMOUNTS (200-400 NL) OF PROTEIN (15 MG/ML IN 20 MM HEPES PH 7.4, 120 MM NACL, 4 MM DTT) AND RESERVOIR SOLUTION (100 MM PHOSPHATE-CITRATE PH 4.2, 200 MM NACL, 50% (V/V) PEG-200) AND EQUILIBRATED AGAINST 80 UL OF RESERVOIR SOLUTION AT 16C. CRYSTALS WERE CRYOPROTECTED BY BRIEF IMMERSION (5-60 S) IN RESERVOIR SOLUTION SUPPLEMENTED WITH 25% GLYCEROL AND WERE RAPIDLY CRYOCOOLED IN LIQUID N2 OR A 100 K STREAM OF N2 GAS.