8UC6

Calpain-7:IST1 Complex

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL14-1
Synchrotron site	SSRL
Beamline	BL14-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-07-27
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	1.19499
Spacegroup name	P 65 2 2
Unit cell lengths	87.841, 87.841, 183.894
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	39.631 - 2.701
R-factor	0.2184
Rwork	0.211
R-free	0.28470
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2kw3
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHASER
Refinement software	PHENIX (1.16)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	39.631	39.630	2.830
High resolution limit [Å]	2.700	8.960	2.700
Rmerge	0.855	0.144	6.810
Rmeas	0.859	0.145	6.843
Rpim	0.080	0.015	0.666
Total number of observations		38774	164526
Number of reflections	12228	417	1581
<I/σ(I)>	11	39.6	1.5
Completeness [%]	100.0	99.1	99.9
Redundancy	114.3	93	104.1
CC(1/2)	0.998	0.999	0.650

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.2	294	Both proteins were purified and concentrated to 20 mg/ml in buffer containing 25 mM Tris, pH 7.2, 150 mM NaCl, 1 mM TCEP, 0.5 mM EDTA. The proteins were mixed in a 1:2 molar ratio (CAPN7:IST1). Crystals grew at 21 C (294 K) in Rigaku Wizard Cryo condition D5 (25% (v/v) 1,2-Porpanediol, 100 mM Sodium phosphate dibasic/Citric acid pH 4.2, 5% (w/v) PEG 3000, 10% (v/v) Glycerol. Crystals were transferred briefly to crystallization buffer supplemented with 25% added glycerol prior to plunging in liquid nitrogen