8HFO

Crystal Structure of Mycobacterium smegmatis MshC in Complex with Compound 7d

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-1
Synchrotron site	ESRF
Beamline	ID23-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-09-24
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.774900
Spacegroup name	P 61
Unit cell lengths	168.113, 168.113, 52.300
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	55.030 - 2.770
R-factor	0.2026
Rwork	0.200
R-free	0.25210
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3c8z
RMSD bond length	0.005
RMSD bond angle	0.772
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.19.2)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	145.590	2.920
High resolution limit [Å]	2.770	2.770
Rmerge	0.098	1.387
Rmeas	0.101	1.421
Rpim	0.023	0.312
Total number of observations	438327	65200
Number of reflections	21805	3164
<I/σ(I)>	17.5	2.6
Completeness [%]	100.0
Redundancy	20.1	20.6
CC(1/2)	0.993	0.898

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	293	Holo protein at 10 mg/mL in 10 mM Tris-HCl pH 7, 100 mM NaCl and 2.5 mM 2-mercaptoethanol was mixed with 1 mM compound 7d and 1 mM TCEP (final concentrations) and incubated on ice for 1 hr. Crystals of MshC in complex with compound 7d were grown in the Morpheus screen by dispensing droplets comprised of 300 nL protein mixture and 150 nL reservoir solution containing PEG550MME 20% (v/v), PEG20000 10% (w/v), Morpheus divalent 60 mM and Morpheus buffer system 3 pH 8.5.