6VER

Human insulin analog: [GluB10,TyrB20]-DOI

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX2
Synchrotron site	Australian Synchrotron
Beamline	MX2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-03-22
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.95364
Spacegroup name	P 21 21 21
Unit cell lengths	26.905, 29.991, 44.864
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	24.933 - 1.047
R-factor	0.188
Rwork	0.186
R-free	0.20510
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Editted version of human insulin component of PDB entry 4OGA
RMSD bond length	0.011
RMSD bond angle	1.274
Data reduction software	XDS
Data scaling software	Aimless (0.5.21)
Phasing software	PHASER
Refinement software	PHENIX ((1.13-2998_1692))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	44.860	1.060
High resolution limit [Å]	1.047	1.047
Rmerge	0.036	0.857
Rmeas	0.040	0.969
Rpim	0.017	0.442
Number of reflections	17679	836
<I/σ(I)>	21.9
Completeness [%]	99.8	98.4
Redundancy	6	4.5
CC(1/2)	0.999	0.735

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		300	Well condition: 100 mM Tri-HCl (pH 8.5) + 0.28 M magnesium formate. Protein: The insulin analog was prepared in a mixture that also contained receptor fragments: 5 mg/ml (IR310.T).Fv83-7 in 10mM HEPES-NaOH (pH7.5) + 0.02% NaN3 plus three mol equivalents of the IR-A alphaCT peptide 704-719 plus 1.8 mol equivalents of the analog. The analog crystallized in isolation from the receptor fragments and it is not known whether the receptor fragments aided crystallization