PDB-4oqw: Crystal structure of mCardinal far-red fluorescent protein

Basic information

• Entry: Database: PDB / ID: 4oqw
• Title: Crystal structure of mCardinal far-red fluorescent protein
• Components: Fluorescent protein FP480Fluorescence
• Keywords: FLUORESCENT PROTEIN
• Function / homology: Green Fluorescent Protein / Green fluorescent protein / Green fluorescent protein-related / Green fluorescent protein / Green fluorescent protein / bioluminescence / Beta Barrel / Mainly Beta / Fluorescent protein FP480
• Biological species: Entacmaea quadricolor (sea anemone)
• Method: X-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 2.21 Å
• Authors: Burg, J.S. / Chu, J. / Lam, A.J. / Lin, M.Z. / Garcia, K.C.
• Citation: Journal: Nat.Methods / Year: 2014; Title: Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein.; Authors: Chu, J. / Haynes, R.D. / Corbel, S.Y. / Li, P. / Gonzalez-Gonzalez, E. / Burg, J.S. / Ataie, N.J. / Lam, A.J. / Cranfill, P.J. / Baird, M.A. / Davidson, M.W. / Ng, H.L. / Garcia, K.C. / Contag, C.H. / Shen, K. / Blau, H.M. / Lin, M.Z.

History

Deposition	Feb 10, 2014	Deposition site: RCSB / Processing site: RCSB
Revision 1.0	Mar 12, 2014	Provider: repository / Type: Initial release
Revision 1.1	May 14, 2014	Group: Database references

Assembly

Deposited unit

A: Fluorescent protein FP480

B: Fluorescent protein FP480

C: Fluorescent protein FP480

D: Fluorescent protein FP480

E: Fluorescent protein FP480

F: Fluorescent protein FP480

G: Fluorescent protein FP480

H: Fluorescent protein FP480

Summary:

• 208 kDa, 8 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	207,989	8
Polymers	207,989	8
Non-polymers	0	0
Water	12,088	671

1

A: Fluorescent protein FP480

Summary:

• defined by author
• 26 kDa, 1 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	25,999	1
Polymers	25,999	1
Non-polymers	0	0
Water	18	1

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

2

B: Fluorescent protein FP480

Summary:

• defined by author
• 26 kDa, 1 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	25,999	1
Polymers	25,999	1
Non-polymers	0	0
Water	18	1

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

3

C: Fluorescent protein FP480

Summary:

• defined by author
• 26 kDa, 1 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	25,999	1
Polymers	25,999	1
Non-polymers	0	0
Water	18	1

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

4

D: Fluorescent protein FP480

Summary:

• defined by author
• 26 kDa, 1 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	25,999	1
Polymers	25,999	1
Non-polymers	0	0
Water	18	1

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

5

E: Fluorescent protein FP480

Summary:

• defined by author&software
• 26 kDa, 1 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	25,999	1
Polymers	25,999	1
Non-polymers	0	0
Water	18	1

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

Calculated values:

Method	PISA

6

F: Fluorescent protein FP480

Summary:

• defined by author
• 26 kDa, 1 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	25,999	1
Polymers	25,999	1
Non-polymers	0	0
Water	18	1

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

7

G: Fluorescent protein FP480

Summary:

• defined by author
• 26 kDa, 1 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	25,999	1
Polymers	25,999	1
Non-polymers	0	0
Water	18	1

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

8

H: Fluorescent protein FP480

Summary:

• defined by author&software
• 26 kDa, 1 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	25,999	1
Polymers	25,999	1
Non-polymers	0	0
Water	18	1

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

Calculated values:

Method	PISA

9

A: Fluorescent protein FP480

C: Fluorescent protein FP480

Summary:

• defined by software
• 52 kDa, 2 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	51,997	2
Polymers	51,997	2
Non-polymers	0	0
Water	36	2

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

Calculated values:

Buried area	4450 Å2
ΔGint	-18 kcal/mol
Surface area	18090 Å2
Method	PISA

10

B: Fluorescent protein FP480

D: Fluorescent protein FP480

Summary:

• defined by software
• 52 kDa, 2 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	51,997	2
Polymers	51,997	2
Non-polymers	0	0
Water	36	2

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1

Calculated values:

Buried area	4050 Å2
ΔGint	-17 kcal/mol
Surface area	18230 Å2
Method	PISA

11

F: Fluorescent protein FP480

G: Fluorescent protein FP480

Summary:

• defined by software
• 52 kDa, 2 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	51,997	2
Polymers	51,997	2
Non-polymers	0	0
Water	36	2

Symmetry operations:

Type	Name	Symmetry operation	Number
identity operation	1_555	x,y,z	1
crystal symmetry operation	2_656	-x+1,y+1/2,-z+1	1

Calculated values:

Buried area	3910 Å2
ΔGint	-15 kcal/mol
Surface area	17680 Å2
Method	PISA

Unit cell

Length a, b, c (Å)	53.200, 136.700, 167.600
Angle α, β, γ (deg.)	90.000, 90.100, 90.000
Int Tables number	4
Space group name H-M	P1211

Components

#1: Protein

A B C D E F G H
Fluorescent protein FP480 / Fluorescence

Details:

Mass: 25998.611 Da / Num. of mol.: 8
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Entacmaea quadricolor (sea anemone) / Production host: Escherichia coli (E. coli) / References: UniProt: D0VX33

#2: Water

ChemComp-HOH / water / Water

Details:

Mass: 18.015 Da / Num. of mol.: 671 / Source method: isolated from a natural source / Formula: H₂O

Experimental details

Experiment

• Experiment: Method: X-RAY DIFFRACTION / Number of used crystals: 1

Sample preparation

• Crystal: Density Matthews: 2.93 ų/Da / Density % sol: 58.02 %
• Crystal grow: Temperature: 300 K / Method: vapor diffusion, hanging drop / pH: 7.6 ; Details: 0.1 M Tris pH 7.6 - 8.0, 0.2 M CaCl2, 10 mM TCEP, 22 - 25% PEG 4000 , VAPOR DIFFUSION, HANGING DROP, temperature 300K

Data collection

• Diffraction: Mean temperature: 100 K
• Diffraction source: Source: SYNCHROTRON / Site: ALS / Beamline: 8.2.2 / Wavelength: 1 Å
• Detector: Type: ADSC QUANTUM 315r / Detector: CCD / Date: Nov 23, 2013
• Radiation: Monochromator: Double crystal, Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
• Radiation wavelength: Wavelength: 1 Å / Relative weight: 1
• Reflection twin: Operator: h,-k,-l / Fraction: 0.5
• Reflection: Resolution: 2.21→47.562 Å / Num. obs: 113638 / % possible obs: 95 % / Observed criterion σ(I): -3 / R_merge(I) obs: 0.131 / Χ²: 1.014 / Net I/σ(I): 5.5

Reflection shell

Resolution (Å)	Rmerge(I) obs	Mean I/σ(I) obs	Num. measured obs	Num. unique obs	Diffraction-ID	% possible all
2.21-2.27	0.822	1.18	15427	8239	1	94.2
2.27-2.33	0.6	1.25	16162	8510	1	98.4
2.33-2.4	0.497	1.49	15693	8243	1	98.6
2.4-2.47	0.426	1.74	15192	7959	1	98.4
2.47-2.55	0.385	1.97	14887	7764	1	98.2
2.55-2.64	0.328	2.4	14241	7450	1	97.9
2.64-2.74	0.264	3.02	13515	7074	1	96.2
2.74-2.85	0.224	3.56	13135	6838	1	96.6
2.85-2.98	0.208	3.87	12555	6558	1	96
2.98-3.13	0.16	4.96	11905	6208	1	95.5
3.13-3.29	0.118	6.67	11048	5785	1	94.2
3.29-3.49	0.086	8.98	10049	5369	1	91.5
3.49-3.74	0.076	9.75	9398	5002	1	91.1
3.74-4.04	0.065	11.16	8617	4577	1	89.2
4.04-4.42	0.05	13.67	7969	4230	1	89.4
4.42-4.94	0.044	14.57	7247	3863	1	90.6
4.94-5.71	0.041	14.52	6752	3480	1	92.2
5.71-6.99	0.049	11.96	5981	2982	1	93.2
6.99-9.88	0.033	14.51	4548	2280	1	91.6
9.88-47.56	0.02	19.57	2461	1227	1	87.8

Phasing

• Phasing: Method: molecular replacement

Processing

Software

Name	Version	Classification	NB
XSCALE		data scaling
PHASER	2.5.5	phasing
PHENIX	1.8.4_1496	refinement
PDB_EXTRACT	3.14	data extraction
XDS		data scaling

Refinement

Method to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.21→47.562 Å / FOM work R set: 0.8347 / σ(F): 1.34 / Phase error: 24.28 / Stereochemistry target values: TWIN_LSQ_F

	Rfactor	Num. reflection	% reflection	Selection details
Rfree	0.2511	2092	1.84 %	RANDOM
Rwork	0.2225	-	-	-
obs	0.2249	113418	94.83 %	-

• Solvent computation: Shrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
• Displacement parameters: B_iso max: 550 Ų / B_iso mean: 42.16 Ų / B_iso min: 4.6 Ų

Refinement step

Cycle: LAST / Resolution: 2.21→47.562 Å

	Protein	Nucleic acid	Ligand	Solvent	Total
Num. atoms	13735	0	0	671	14406

Refine LS restraints

Refine-ID	Type	Dev ideal	Number
X-RAY DIFFRACTION	f_bond_d	0.004	14044
X-RAY DIFFRACTION	f_angle_d	0.861	18964
X-RAY DIFFRACTION	f_chiral_restr	0.035	2026
X-RAY DIFFRACTION	f_plane_restr	0.003	2417
X-RAY DIFFRACTION	f_dihedral_angle_d	13.26	5130

LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 14

Resolution (Å)	Rfactor Rfree	Num. reflection Rfree	Rfactor Rwork	Num. reflection Rwork	Num. reflection all	% reflection obs (%)
2.2111-2.2664	0.3534	137	0.3053	7659	7796	91
2.2664-2.3276	0.3369	155	0.2728	8240	8395	96
2.3276-2.3961	0.2463	147	0.2612	8240	8387	97
2.3961-2.4734	0.2886	144	0.2599	8205	8349	97
2.4734-2.5617	0.2989	149	0.2501	8203	8352	96
2.5617-2.6643	0.3001	142	0.2485	8209	8351	96
2.6643-2.7854	0.2442	141	0.2405	8064	8205	95
2.7854-2.9322	0.2717	143	0.2315	8039	8182	94
2.9322-3.1157	0.2617	148	0.2273	8002	8150	94
3.1157-3.3559	0.2508	141	0.2153	7843	7984	92
3.3559-3.6931	0.2353	136	0.2139	7623	7759	90
3.6931-4.2262	0.2278	136	0.1962	7534	7670	88
4.2262-5.3195	0.2057	137	0.1703	7639	7776	89
5.3195-29.6798	0.2662	145	0.235	7820	7965	90

Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

ID	L11 (°2)	L12 (°2)	L13 (°2)	L22 (°2)	L23 (°2)	L33 (°2)	S11 (Å °)	S12 (Å °)	S13 (Å °)	S21 (Å °)	S22 (Å °)	S23 (Å °)	S31 (Å °)	S32 (Å °)	S33 (Å °)	T11 (Å2)	T12 (Å2)	T13 (Å2)	T22 (Å2)	T23 (Å2)	T33 (Å2)	Origin x (Å)	Origin y (Å)	Origin z (Å)
1	0.0405	-0.1588	-0.0999	1.2349	0.2806	0.2399	-0.0816	-0.0693	0.1407	0.288	0.0635	-0.0214	0.0258	0.0704	0.0275	0.0629	0.0098	0.0016	0.2027	0.0017	0.1246	19.6119	-22.5762	176
2	0.8316	-0.6792	0.0733	2.4062	0.4011	0.1919	0.0658	-0.0634	0.1302	0.0114	-0.0286	0.1432	0.0937	-0.1775	0.0117	0.1011	-0.0387	0.0096	0.1174	0.0312	0.065	13.7944	-21.0695	172.5938
3	0.2095	-0.0015	-0.1324	0.2081	0.1158	0.1878	-0.0804	-0.1015	-0.0279	0.0075	0.1038	0.0977	-0.0018	-0.008	0.0324	0.0999	-0.0122	0.0024	0.0916	0.0331	0.1033	23.5237	-23.0408	168.9968
4	0.5516	-0.3875	0.4171	1.5259	-0.849	1.2408	0.0418	-0.0873	0.0345	0.1084	-0.0035	0.0957	-0.0685	-0.0475	0.0135	0.0291	0.0097	0.0201	0.088	-0.0118	0.0751	21.9933	-12.0485	168.2731
5	2.0824	-0.8564	0.4813	1.0958	-0.0837	0.8131	-0.0556	-0.117	0.1193	0.0798	-0.0015	-0.1152	-0.0862	-0.1809	-0.0168	0.1081	-0.0107	0.0196	0.1067	0.0185	0.0217	24.9496	-19.2432	158.9542
6	0.0988	-0.079	0.069	0.0838	-0.0066	0.1277	0.0575	0.0944	0.1054	0.0093	-0.0098	-0.1281	0.005	-0.0004	0.0896	0.0511	0.059	-0.0384	0.0842	0.0298	0.0616	25.4193	-15.9046	158.1412
7	0.2469	-0.155	-0.4449	0.9248	0.4667	1.0362	0.0805	-0.0582	-0.0677	-0.0778	0.0345	-0.1537	0.0775	0.0333	-0.0829	0.1085	0.0135	0.0384	0.1206	0.0322	0.0709	27.6781	-25.7822	167.424
8	2.4044	-0.9334	-0.7087	0.9921	0.4886	0.6772	0.1211	0.1441	-0.116	-0.1739	-0.1368	-0.0673	-0.0683	-0.0159	0.0358	0.1207	-0.0203	-0.014	0.1033	0.0528	0.0809	22.4928	-26.8827	160.227
9	0.561	-0.289	-0.0093	1.1158	-0.3677	0.3222	0.0612	-0.1298	0.019	0.2674	0.0112	-0.0017	-0.0209	0.0536	0.007	0.2426	0.016	-0.0554	0.1625	-0.0068	0.0168	30.4189	10.2333	179.0409
10	0.9558	0.2702	-0.0897	0.5107	-0.5823	0.7237	-0.0193	0.0134	-0.233	-0.0358	-0.0036	-0.0268	0.1583	-0.077	-0.0006	0.2154	0.0081	0.0117	0.0876	-0.0387	0.0968	22.3547	19.2591	168.8341
11	0.392	-0.0615	-0.155	0.7006	-0.1566	0.4699	0.0807	-0.1223	0.0452	0.0228	-0.0413	0.054	0.0816	-0.0131	-0.0073	0.1613	-0.012	-0.043	0.1408	-0.0046	0.001	29.9852	7.8735	167.2338
12	0.5747	0.1731	-0.2365	1.0104	-0.1475	0.7179	0.0018	0.0113	0.0133	-0.0166	-0.1606	-0.1445	0.0573	0.1153	0.0936	0.1279	-0.0434	-0.0116	0.1296	-0.009	0.103	35.1239	16.9921	168.5628
13	0.6983	0.4542	0.3139	2.6159	1.4335	1.6059	-0.0593	0.3017	0.0396	-0.2402	0.0688	-0.0615	0.0162	-0.0479	-0.0531	0.155	0.0389	0.0211	0.3359	-0.0247	0.1029	42.3385	-20.4414	132.2684
14	0.0854	-0.0215	0.1709	0.1548	-0.0901	0.3561	0.0836	0.1575	0.0424	-0.1546	-0.0149	-0.1926	0.0962	0.1136	0.0202	0.1465	0.028	0.0553	0.2149	-0.0026	0.1801	46.1775	-22.5862	134.9282
15	1.0082	0.5058	0.124	1.1195	0.226	0.4186	0.1118	0.069	-0.0243	-0.024	-0.0918	-0.1379	0.0455	-0.1283	-0.0089	0.136	-0.0235	-0.0198	0.2187	-0.0282	0.1133	36.2333	-22.6584	138.2377
16	0.4134	0.0087	0.0835	0.2747	0.1954	0.5197	-0.0046	0.1377	0.165	-0.0334	-0.0778	0.0083	0.001	-0.0473	0.0276	0.0257	-0.009	0.0014	0.2224	-0.0281	0.1496	35.8926	-11.872	144.8041
17	0.119	0.0115	-0.0129	0.3651	0.1972	0.2924	0.0285	0.1831	-0.0016	-0.1268	-0.0183	0.0124	0.0462	0.0063	0.0185	0.1481	0.0257	0.0115	0.3173	0.024	0.0397	41.8755	-12.6456	138.4392
18	1.1452	0.329	0.1231	0.3173	0.3073	0.3419	-0.0591	0.1288	0.0502	-0.0413	-0.0065	-0.0017	-0.0491	0.1648	-0.065	0.089	0.009	-0.0085	0.1842	0.0025	0.0003	35.5607	-19.746	148.5367
19	0.4297	-0.0744	0.5153	0.1416	-0.0977	1.017	0.0274	0.033	0.0089	-0.0074	-0.0571	-0.0173	0.0942	0.0312	0.0042	0.0931	-0.0504	0.0281	0.1906	0.0099	0.004	35.7745	-16.5612	149.8779
20	0.8461	0.1785	-0.1269	0.5109	0.0193	0.5254	0.0103	0.0369	0.0183	-0.1279	0.0108	0.0618	-0.0587	0.0204	0.0509	0.2409	-0.0037	-0.0078	0.2846	0.0039	0.0063	31.6192	-24.6498	139.5526
21	0.9699	0.4744	-0.4521	1.0562	-0.2408	0.9311	-0.0186	0.1346	-0.0104	0.054	0.0188	0.0955	-0.0076	0.0159	-0.0501	0.1773	-0.0163	-0.0061	0.2299	-0.0066	0.0245	36.2494	-27.6992	146.2797
22	0.2725	0.162	-0.2771	0.4189	-0.1829	0.8163	0.0606	0.0236	0.0101	-0.0232	0.0096	-0.0567	-0.179	0.2183	0.2154	0.2071	-0.0189	-0.0113	0.3509	0.0375	0.0673	46.1258	16.8813	133.5607
23	0.3926	0.0494	-0.0901	0.1771	0.0251	0.4725	0.1299	0.18	0.0173	-0.1133	-0.0202	0.0017	-0.0447	0.0484	0.0974	0.1517	0.0136	-0.0207	0.2746	-0.0433	-0.1385	36.5272	9.4802	140.7415
24	0.1363	-0.1742	-0.0941	0.5484	0.3923	1.613	0.0377	0.0645	0.0649	-0.0065	-0.0358	-0.0157	-0.1718	0.0153	0.0186	0.1901	-0.0096	-0.0038	0.2283	0.0081	0.014	38.7385	17.2954	144.7038
25	1.3211	0.2888	-0.3576	0.1913	0.3459	1.5686	-0.0687	-0.007	0.0266	0.0541	-0.0056	-0.0402	0.1167	0.1122	0.0255	0.4157	-0.0486	-0.0609	0.4463	0.0385	0.2224	14.9691	-3.5036	119.4773
26	5.2693	0.4558	-0.0434	2.3427	-1.1584	5.0922	-0.0422	0.3616	-0.4406	-0.3131	0.0358	-0.4913	0.5193	0.4594	-0.0102	0.6101	-0.0309	0.0625	0.5332	0.1064	0.5991	6.7268	2.8442	115.4622
27	1.9471	0.6112	-0.146	2.12	-0.0632	1.0242	-0.0067	-0.1927	0.1948	0.1216	0.0065	0.0181	-0.064	-0.0453	0.0305	0.6041	-0.042	-0.1322	0.4735	0.1269	0.4549	24.9941	-9.5088	106.4152
28	0.4509	-0.5475	0.2565	0.6629	-0.3122	0.1904	-0.013	0.1586	-0.0271	0.1309	0.0043	-0.1097	0.079	0.0699	-0.0124	0.4689	-0.0596	-0.0647	0.3931	-0.0219	0.3975	11.716	4.6569	110.0894
29	0.1153	0.1217	0.1997	0.4672	0.0296	0.4376	0.1399	0.1211	-0.0691	0.1832	0.1158	0.0788	-0.1686	0.1062	-0.1012	0.3896	-0.0843	0.0185	0.3785	-0.0026	0.5022	12.5122	-3.3949	105.2776
30	0.1245	0.0455	-0.0479	2.0219	0.4304	0.8079	0.0705	-0.1644	-0.112	0.1313	-0.0879	0.1015	-0.0736	0.0024	-0.0128	0.4712	-0.0685	-0.0614	0.5431	-0.0246	0.3349	21.1836	31.945	117.4826
31	1.2892	-0.0335	2.0756	2.4731	-0.9603	6.2654	-0.0451	-0.1724	0.0118	0.2357	0.1075	0.041	-0.2689	-0.2425	0.0385	0.3026	-0.1259	-0.0174	0.5812	0.0661	0.1505	26.2059	24.3535	107.8973
32	0.0083	-0.0041	0.0046	0.0117	0.0014	0.0034	-0.0198	0.2687	-0.028	-0.4872	-0.0335	-0.4706	0.2868	0.4573	0.0189	0.5437	0.0297	0.0856	0.5208	-0.0343	0.4387	7.0351	36.2768	116.4635
33	0.3696	0.1593	0.2163	0.4985	-0.2224	0.502	0.1046	-0.1	-0.07	0.0553	-0.1098	-0.1787	0.1098	-0.2357	0.0222	0.39	-0.1068	-0.0386	0.5482	0.0269	0.2993	12.8429	27.6837	107.7719
34	0.0961	0.1103	-0.1904	0.6782	0.0974	0.5859	0.0099	0.1342	0.049	-0.1185	-0.0128	-0.0624	-0.0078	0.021	-0.0049	0.5982	-0.0755	0.0237	0.6203	-0.0456	0.2413	17.5122	37.0797	107.1014
35	0.0471	0.0221	-0.0718	0.068	-0.0565	0.1124	0.1308	0.1736	0.0273	-0.0051	0.0177	-0.0717	-0.0731	-0.071	-0.0802	0.5259	-0.0128	0.0079	0.59	-0.0569	0.2873	59.2478	-35.6516	87.8748
36	0.5663	0.0093	0.3599	0.1856	-0.4841	1.521	-0.0031	0.0022	-0.0769	0.1539	0.1038	-0.0023	-0.043	0.0443	-0.0147	0.4433	0.0852	-0.1305	0.5087	-0.0529	0.4473	48.7535	-37.8252	87.4841
37	0.6689	-0.3482	-0.6814	0.2439	0.3612	0.6968	0.0557	-0.0169	-0.1524	0.0973	-0.1204	0.0162	-0.1589	-0.0934	-0.0097	0.3749	0.0998	-0.0157	0.5341	-0.0662	0.411	56.7818	-43.0798	75.9336
38	0.5127	-0.0962	-0.3062	0.2523	0.1467	0.3717	0.0097	-0.1612	0.0877	-0.0124	0.0543	-0.0655	-0.1393	-0.0798	-0.0236	0.4924	0.023	-0.0673	0.5264	-0.0194	0.2474	51.5783	-34.1423	80.7854
39	0.8793	0.4563	0.3693	0.5443	0.6391	0.8055	0.0417	0.0153	0.0414	-0.0435	0.0183	0.0915	0.0368	-0.0362	-0.0226	0.3432	-0.0287	-0.0372	0.5047	-0.0655	0.6097	47.4688	-7.0107	71.4869
40	0.0068	0.0083	0.0032	0.0084	0.0034	0.0013	-0.0641	-0.0419	0.0949	0.0152	0.1013	0.3494	-0.0341	-0.098	-0.0918	0.5964	-0.1343	0.1318	0.6334	-0.0573	0.4388	55.4423	-3.4239	70.5977
41	0.5641	0.7204	0.004	0.9219	0.0065	0.0002	-0.0099	0.1844	0.1756	-0.0059	-0.0029	0.1308	-0.0328	0.0275	-0.0318	0.8061	0.0154	-0.059	0.6073	0.0064	0.363	55.8131	-1.0561	78.7047
42	1.8711	0.1918	1	0.368	0.2792	1.1826	0.0535	0.1029	0.0847	-0.1132	-0.0842	0.2904	-0.0535	-0.0029	0.0292	0.4682	-0.0081	-0.1322	0.5287	-0.0155	0.2657	47.3534	-0.5035	83.731
43	0.1972	0.1686	0.2309	0.1802	0.1634	0.293	-0.1167	-0.0305	0.1401	-0.1208	0.0184	-0.0248	-0.0653	-0.112	0.0012	0.5669	0.0465	0.0774	0.5554	-0.0508	0.3191	57.1479	2.6135	84.0086
44	0.0566	0	0.1214	0.5625	-0.1525	0.3005	-0.0116	0.1874	-0.0702	-0.117	-0.0558	0.203	-0.0573	0.1996	0.0601	0.4126	0.0138	-0.1327	0.5438	0.0512	0.3872	52.481	-1.3544	84.3877
45	0.003	0.0049	-0.0024	0.0063	-0.0038	0.0017	-0.0333	-0.1532	-0.1218	0.1049	-0.1202	-0.2193	0.1337	0.3704	0.2191	0.6256	0.0208	0.0705	0.3298	0.0136	0.51	57.5827	-9.4553	85.3365

Refinement TLS group

ID	Refine-ID	Refine TLS-ID	Selection details	Auth asym-ID	Auth seq-ID
1	X-RAY DIFFRACTION	1	chain 'A' and (resid 1 through 21 )	A	0
2	X-RAY DIFFRACTION	2	chain 'A' and (resid 22 through 47 )	A	0
3	X-RAY DIFFRACTION	3	chain 'A' and (resid 48 through 87 )	A	0
4	X-RAY DIFFRACTION	4	chain 'A' and (resid 88 through 134 )	A	0
5	X-RAY DIFFRACTION	5	chain 'A' and (resid 135 through 152 )	A	0
6	X-RAY DIFFRACTION	6	chain 'A' and (resid 153 through 179 )	A	0
7	X-RAY DIFFRACTION	7	chain 'A' and (resid 180 through 205 )	A	0
8	X-RAY DIFFRACTION	8	chain 'A' and (resid 206 through 228 )	A	0
9	X-RAY DIFFRACTION	9	chain 'B' and (resid 1 through 61 )	B	0
10	X-RAY DIFFRACTION	10	chain 'B' and (resid 62 through 87 )	B	0
11	X-RAY DIFFRACTION	11	chain 'B' and (resid 88 through 205 )	B	0
12	X-RAY DIFFRACTION	12	chain 'B' and (resid 206 through 226 )	B	0
13	X-RAY DIFFRACTION	13	chain 'C' and (resid 4 through 21 )	C	0
14	X-RAY DIFFRACTION	14	chain 'C' and (resid 22 through 47 )	C	0
15	X-RAY DIFFRACTION	15	chain 'C' and (resid 48 through 87 )	C	0
16	X-RAY DIFFRACTION	16	chain 'C' and (resid 88 through 100 )	C	0
17	X-RAY DIFFRACTION	17	chain 'C' and (resid 101 through 134 )	C	0
18	X-RAY DIFFRACTION	18	chain 'C' and (resid 135 through 152 )	C	0
19	X-RAY DIFFRACTION	19	chain 'C' and (resid 153 through 179 )	C	0
20	X-RAY DIFFRACTION	20	chain 'C' and (resid 180 through 205 )	C	0
21	X-RAY DIFFRACTION	21	chain 'C' and (resid 206 through 228 )	C	0
22	X-RAY DIFFRACTION	22	chain 'D' and (resid -1 through 19 )	D	0
23	X-RAY DIFFRACTION	23	chain 'D' and (resid 20 through 179 )	D	0
24	X-RAY DIFFRACTION	24	chain 'D' and (resid 180 through 226 )	D	0
25	X-RAY DIFFRACTION	25	chain 'E' and (resid 2 through 47 )	E	0
26	X-RAY DIFFRACTION	26	chain 'E' and (resid 48 through 66 )	E	0
27	X-RAY DIFFRACTION	27	chain 'E' and (resid 67 through 87 )	E	0
28	X-RAY DIFFRACTION	28	chain 'E' and (resid 88 through 153 )	E	0
29	X-RAY DIFFRACTION	29	chain 'E' and (resid 154 through 224 )	E	0
30	X-RAY DIFFRACTION	30	chain 'F' and (resid 5 through 47 )	F	0
31	X-RAY DIFFRACTION	31	chain 'F' and (resid 48 through 61 )	F	0
32	X-RAY DIFFRACTION	32	chain 'F' and (resid 62 through 87 )	F	0
33	X-RAY DIFFRACTION	33	chain 'F' and (resid 88 through 205 )	F	0
34	X-RAY DIFFRACTION	34	chain 'F' and (resid 206 through 224 )	F	0
35	X-RAY DIFFRACTION	35	chain 'G' and (resid 2 through 74 )	G	0
36	X-RAY DIFFRACTION	36	chain 'G' and (resid 75 through 113 )	G	0
37	X-RAY DIFFRACTION	37	chain 'G' and (resid 114 through 169 )	G	0
38	X-RAY DIFFRACTION	38	chain 'G' and (resid 170 through 226 )	G	0
39	X-RAY DIFFRACTION	39	chain 'H' and (resid 1 through 18 )	H	0
40	X-RAY DIFFRACTION	40	chain 'H' and (resid 24 through 47 )	H	0
41	X-RAY DIFFRACTION	41	chain 'H' and (resid 48 through 73 )	H	0
42	X-RAY DIFFRACTION	42	chain 'H' and (resid 74 through 108 )	H	0
43	X-RAY DIFFRACTION	43	chain 'H' and (resid 109 through 169 )	H	0
44	X-RAY DIFFRACTION	44	chain 'H' and (resid 170 through 209 )	H	0
45	X-RAY DIFFRACTION	45	chain 'H' and (resid 210 through 225 )	H	0