9Q39

Structure of a sortase-linked cytochrome c peroxidase - cytochrome c fusion protein

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-E
Synchrotron site	APS
Beamline	24-ID-E
Temperature [K]	100
Detector technology	PIXEL
Collection date	2025-03-27
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.9792
Spacegroup name	C 1 2 1
Unit cell lengths	160.658, 46.409, 60.604
Unit cell angles	90.00, 107.40, 90.00

Refinement procedure

Resolution	44.420 - 3.240
R-factor	0.2817
Rwork	0.276
R-free	0.33130
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.004
RMSD bond angle	0.730
Data reduction software	RAPD
Data scaling software	RAPD
Phasing software	PHASER
Refinement software	PHENIX (1.21.2_5419)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	44.420	3.490
High resolution limit [Å]	3.240	3.240
Rmerge	0.257	0.626
Rmeas	0.315	0.766
Rpim	0.179	0.434
Number of reflections	6454	1337
<I/σ(I)>	4.4
Completeness [%]	94.4	95.7
Redundancy	2.7
CC(1/2)	0.967	0.679

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.5	295	Upon purification, CcP-GGSG(2x)LPATGGG was buffer exchanged into filtered nanopure water and concentrated to 0.5 mM. Initial crystal hits were obtained using a Gryphon robot (Arts Robbins Instrument). Larger crystals were optimized via vapor diffusion in 4 microliter sitting drops, which were mixed 1:1 with well solution (14-20 % PEG 8000, 40 mM KH2PO4, 20 % glycerol, pH 3.0-5.5). Crystals growing under these conditions formed within a week.