7QSW

L8S8-complex forming RubisCO derived from ancestral sequence reconstruction of the last common ancestor of SSU-bearing 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	P 43 21 2
Unit cell lengths	131.873, 131.873, 305.463
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.960 - 1.800
R-factor	0.1408
Rwork	0.141
R-free	0.15920
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]	19.963	19.963	1.900
High resolution limit [Å]	1.800	5.690	1.800
Rmerge		0.050	1.015
Rmeas	0.128	0.052	1.054
Rpim	0.034	0.014	0.282
Total number of observations	3394733	108989	492040
Number of reflections	247678	8256	35779
<I/σ(I)>	15.3	43	2.8
Completeness [%]	99.9	97.7	100
Redundancy	13.7	13.2	13.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	9.4	288	Purified enzyme (1.5 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 0.125 mM CABP and 2 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 9.4. Before flash freezing the crystals in liquid nitrogen PEG200 was added to the mother liquor as cryoprotectant to a final concentration of 33 % (w/v).