8QMW

Non-obligately L8S8-complex forming RubisCO derived from ancestral sequence reconstruction and rational engineering in L8S8 complex with substitutions R269W, E271R, L273N

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P14 (MX2)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P14 (MX2)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-12-02
Detector	DECTRIS EIGER2 X CdTe 16M
Wavelength(s)	0.6888
Spacegroup name	C 1 2 1
Unit cell lengths	206.010, 106.440, 108.490
Unit cell angles	90.00, 113.09, 90.00

Refinement procedure

Resolution	24.880 - 1.750
R-factor	0.1451
Rwork	0.145
R-free	0.17450
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.013
RMSD bond angle	1.148
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX ((1.20.1_4487: ???))

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	24.880	24.880	1.800
High resolution limit [Å]	1.750	7.830	1.750
Rmerge	0.124	0.051	0.673
Rmeas	0.133	0.055	0.724
Number of reflections	213396	2392	15647
<I/σ(I)>	11.11
Completeness [%]	98.5
Redundancy	7.2
CC(1/2)	0.997	0.998	0.820

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	9.1	289	Purified enzyme (9 mg/mL) in 25 mM Tricine-NaOH, 75 mM NaCl, pH 8.0 was incubated at 3% (v/v) CO2 in air and 30 degrees C for 1 hour in the presence of 0.35 mM CABP and 5.6 mM MgCl2. The enzyme was then mixed in a 1:1 ratio with 0.2 M BIS-TRIS propane, 20 % (w/v) polyethylene glycol 4000, pH 9.1. Drops were supplemented with 25 % (v/v) PEG200 before flash freezing crystals in lquid nitrogen.