7QSV

L8-complex forming RubisCO derived from ancestral sequence reconstruction of the last common ancestor of Form I'' and Form I RubisCOs

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	2021-08-20
Detector	DECTRIS EIGER2 X 16M
Wavelength(s)	0.97626
Spacegroup name	C 1 2 1
Unit cell lengths	114.007, 203.445, 174.974
Unit cell angles	90.00, 100.43, 90.00

Refinement procedure

Resolution	29.860 - 2.100
R-factor	0.189
Rwork	0.189
R-free	0.22300
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]	29.863	29.863	2.210
High resolution limit [Å]	2.100	6.640	2.100
Rmerge		0.044	0.911
Rmeas	0.146	0.047	0.983
Rpim	0.054	0.017	0.365
Total number of observations	1613528	51877	235574
Number of reflections	225493	7198	32927
<I/σ(I)>	8.9	27.2	2.1
Completeness [%]	99.2	98.6	99.3
Redundancy	7.2	7.2	7.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	288	Purified enzyme (7.5 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 0.25 mM CABP and 4 mM MgCl2. The enzyme was then mixed in a 1:1 ratio with 40 % v/v Glycerol ethoxylate. Crystals were flash frozen in liquid nitrogen.