7QSY

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

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	2021-07-09
Detector	DECTRIS PILATUS3 2M
Wavelength(s)	0.873128
Spacegroup name	I 1 2 1
Unit cell lengths	109.352, 106.608, 192.289
Unit cell angles	90.00, 98.74, 90.00

Refinement procedure

Resolution	29.340 - 2.100
R-factor	0.1708
Rwork	0.170
R-free	0.19690
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]	29.829	29.829	2.210
High resolution limit [Å]	2.100	6.640	2.100
Rmerge		0.088	0.607
Rmeas	0.244	0.096	0.656
Rpim	0.092	0.036	0.245
Total number of observations	856910	24308	127353
Number of reflections	124497	3610	18077
<I/σ(I)>	6.3	13.7	2.6
Completeness [%]	98.1	87.3	97.8
Redundancy	6.9	6.7	7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.7	288	Purified enzyme (5 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 0.2 mM CABP and 3.2 mM MgCl2. The enzyme was then mixed in a 1:1 ratio with 0.2 M Hepes, 20 % (w/v) polyethylene glycol 4000, pH 7.7. Before flash freezing the crystals in liquid nitrogen PEG200 was added to the mother liquor as cryoprotectant to a final concentration of 40 % (w/v).