Combined protein construct and synthetic gene engineering for heterologous protein expression and crystallization using Gene Composer

Table 3 Crystallographic data for Bacillus subtilis FtsZ structures.

Data Collection	2RHH	2RHJ	2RHL	2RHO
Unit Cell a, b, c (Å)	66.37, 66.37 152.09	74.14, 82.06, 117.51	82.29, 97.58, 135.45	82.30, 97.16, 134.72
Space group	P4₁2₁2	C222₁	P2₁2₁2₁	P2₁2₁2₁
Resolution (Å) range/highest resolution shell	50-2.0/(2.07-2.0)	50-1.76/(1.82-1.76)	30-2.45/(2.54-2.45)	50-2.45/(2.54-2.45)
Wavelength (Å)	0.97934	0.97934	0.97934	0.97934
Observed Reflections	267,428	236,328	262,233	228,703
Unique Reflections	23,413	34,949	41,246	40,114
Average I/(sigI)^a	19.8 (1.4)	27.4 (2.1)	29.0 (2.6)	24.7 (2.0)
R_sym (%)^a,c	8.7 (44.9)	7.1 (64.2)	6.9 (47.6)	5.0 (56.8)
Completeness (%)^a	98.3 (88.6)	98.0 (88.8)	99.2 (98.2)	98.1 (94.9)
Redundancy^a	11.4 (6.0)	6.8 (4.6)	6.4 (5.5)	5.7 (4.7)
Refinement
Resolution (Å)	50 - 2.0	50 – 1.76	30 - 2.45	50 - 2.45
Reflections (working/test)	22,153/1,194	33,173/1,752	38,318/2,039	37,673/2,005
R_cryst/R_free (%)^d	21.2/25.7	18.0/20.8	22.7/27.0	22.7/27.0
Number of atoms (protein/ligand/water)	2191/5/114	2279/64/172	2199, 2290^b/ 28/97	2199, 2178^b/32, 28/99
r.m.s.d. bond lengths (Å)	0.019	0.017	0.015	0.014
r.m.s.d. bond angles (°)	1.580	1.520	1.521	1.508
Average B factor all (Å²)	34.2	27.3	67.8	54.8
Average B factor protein (Å²)	36.8	25.7	68.0/67.8^b	55.2/54.7^b
Average B factor ligand (Å²)	31.7	43.6	59.1/53.4^b	44.4/43.1^b
Average B factor water (Å²)	37.1	39.5	67.7	52.4
Coordinate Error (Å)^e	0.170	0.104	0.235	0.235
Ramachandran Analysis (%)
Most Favored	95.8	96.6	94.8/95.4^b	96.5/96.1^b
Additionally Allowed	4.2	3.4	5.2/4.6^b	3.5/3.5^b

^aValues in parenthesis are for the highest resolution shells.
^bValues for chains A and B respectively when applicable.
^cR_sym = (Σ_hklΣ_i|I_i(hkl) - <I(hkl)>|/Σ_hklΣ_i I_i(hkl), I_i(hkl) is the intensity measured for the i^threflection and <I(hkl)> is the mean intensity of reflections with indices hkl.
^d R_cryst = (Σ_hkl || F_obs(hkl)| - |F_calc(hkl)||)/Σ_hkl| F_obs(hkl)|; R_free is calculated in an identical manner using 5% of randomly selected reflections.
^eEstimated standard uncertainty based on R_free.

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