7QSZ

Non-obligately L8S8-complex forming RubisCO derived from ancestral sequence reconstruction and rational engineering in L8 complex with substitution e170N

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-2
Synchrotron site	ESRF
Beamline	ID23-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-12-02
Detector	DECTRIS PILATUS3 2M
Wavelength(s)	0.873128
Spacegroup name	C 2 2 21
Unit cell lengths	122.434, 204.691, 148.457
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.330 - 2.250
R-factor	0.1616
Rwork	0.161
R-free	0.19390
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6ura
Data reduction software	XDS (20210323)
Data scaling software	SCALA (3.3.22)
Phasing software	PHENIX (1.18.2_3874)
Refinement software	PHENIX (1.18.2_3874)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	29.330	29.325	2.370
High resolution limit [Å]	2.250	7.120	2.250
Rmerge		0.060	1.176
Rmeas	0.269	0.062	1.216
Rpim	0.069	0.017	0.309
Total number of observations		39373	196536
Number of reflections	88310	2923	12772
<I/σ(I)>	10.6	33.3	2.6
Completeness [%]	99.9	98.3	100
Redundancy	15	13.5	15.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	288	Purified enzyme (2.25 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 h in the presence of 0.182 mM CABP and 2.9 mM MgCl2. The enzyme was then mixed in a 1:1 ratio with 0.1 M Hepes, 0.2 M MgCl2, 30 % (w/v) polyethylene glycol 400, pH 7.5. Crystals were flash frozen in liquid nitrogen (no additional cryoprotectant added).