9YPL

MboA with Leu-Ala-Arg peptide substrate bound

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS-II BEAMLINE 17-ID-1
Synchrotron site	NSLS-II
Beamline	17-ID-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2025-03-28
Detector	DECTRIS EIGER X 9M
Wavelength(s)	0.92
Spacegroup name	P 43 21 2
Unit cell lengths	157.741, 157.741, 157.014
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	43.750 - 2.200
R-factor	0.172
Rwork	0.171
R-free	0.19360
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.005
RMSD bond angle	0.703
Data reduction software	autoPROC (1.0.5)
Data scaling software	Aimless (0.8.2)
Phasing software	PHASER (2.8.3)
Refinement software	PHENIX (2.0_5761)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	43.750	2.240
High resolution limit [Å]	2.200	2.200
Rmerge	0.257	2.284
Rmeas	0.266	2.366
Rpim	0.070	0.617
Number of reflections	100535	4924
<I/σ(I)>	13.5	1.8
Completeness [%]	100.0	100
Redundancy	27.64	28.34
CC(1/2)	0.998	0.634

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.8	298	MboA crystals were prepared via the hanging drop vapor diffusion method. Hanging drops were prepared by combining equal volumes of MboA (9 mg per mL) protein solution and reservoir solution (0.4 M lithium sulfate and 0.1 M sodium acetate, pH 4.8) for a total drop volume of 2 uL. The substrate bound structure was prepared by incubating apo crystals with 0.3 uL of 100 mM LAR in 0.1 M sodium acetate, pH 4.6 for two hours prior to looping. Crystals were cryoprotected with the addition of 0.75 uL of 40% glycerol in 0.1 M sodium acetate, pH 4.6 and flash frozen in LN2