9FWD

Structure of indole-3-acetic acid-amido synthetase GH3.6 from A.thaliana in complex with AMP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SOLEIL BEAMLINE PROXIMA 2
Synchrotron site	SOLEIL
Beamline	PROXIMA 2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-05-11
Detector	DECTRIS EIGER X 9M
Wavelength(s)	0.987
Spacegroup name	P 64
Unit cell lengths	196.975, 196.975, 65.206
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	28.040 - 1.925
R-factor	0.2046
Rwork	0.203
R-free	0.24020
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.008
RMSD bond angle	1.010
Data reduction software	XDS (Jun 30, 2023)
Data scaling software	Aimless (0.7.7)
Phasing software	PHASER
Refinement software	BUSTER (2.10.4)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	85.293	85.293	2.178
High resolution limit [Å]	1.925	6.545	1.925
Rmerge	0.174	0.043	1.727
Rmeas	0.179	0.044	1.776
Rpim	0.039	0.010	0.411
Total number of observations	1224027	58960	50777
Number of reflections	58596	2927	2932
<I/σ(I)>	14.56	51.4	2
Completeness [%]	95.4	100	76.6
Completeness (spherical) [%]	53.5	100.0	8.7
Completeness (ellipsoidal) [%]	95.4	100.0	76.6
Redundancy	20.89	20.14	17.32
CC(1/2)	0.998	0.999	0.637
Anomalous completeness (spherical)	53.5	99.7	9.0
Anomalous completeness	95.3	99.7	76.6
Anomalous redundancy	10.7	10.8	8.7
CC(ano)	-0.199	-0.528	-0.032
|DANO|/σ(DANO)	0.8	0.6	0.8

Diffraction limits	Principal axes of ellipsoid fitted to diffraction cut-off surface
2.613 Å	1.000, 1.000, 1.000
2.613 Å	0.000, 0.000, 0.000
1.858 Å	0.000, 0.000, 0.000
Criteria used in determination of diffraction limits	local <I/sigmaI> ≥ 1.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	293	0.1 M MES pH 6.5, 20 % PEG 4000, 0.6 M NaCl; protein at final concentration of 8.2 mg/ml and 10 mM AMP