5TD4

Starch binding sites on the Human pancreatic alpha amylase D300N variant complexed with an octaose substrate.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL7-1
Synchrotron site	SSRL
Beamline	BL7-1
Temperature [K]	100
Detector technology	CCD
Collection date	2008-05-01
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.976067
Spacegroup name	P 21 21 21
Unit cell lengths	52.210, 73.640, 135.380
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	38.477 - 2.300
R-factor	0.1531
Rwork	0.151
R-free	0.18860
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4x9y
RMSD bond length	0.009
RMSD bond angle	1.109
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX (1.10.1_2155)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	38.464	2.360
High resolution limit [Å]	2.299	2.300
Rmerge	0.048	0.220
Number of reflections	21229
<I/σ(I)>	24.23	8.13
Completeness [%]	88.8	99.7
Redundancy	4.4	4.5
CC(1/2)	0.999	0.975

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	EVAPORATION	7.5	293	MPD 50-58%, 0.1M sodium cacodylate. After crystal reached full size they were soaked in 200mM octaose for 24h prior to data collection, and then infused with a further aliquot of octaose just before freezing the complexed crystal in liquid nitrogen for use in X-ray diffraction analyses.