PDB-8ooq: Glutamine synthetase from Methanothermococcus thermolithotrophicu...

Basic information

• Entry: Database: PDB / ID: 8ooq
• Title: Glutamine synthetase from Methanothermococcus thermolithotrophicus in complex with 2-oxoglutarate and Mg at 2.91 A resolution
• Components: Glutamine synthetase from Methanothermococcus thermolithotrophicus
• Keywords: LIGASE / Nitrogen-assimilation / methanogenic archaea / allosteric activation / hydrogenotrophic / thermophile / marine / 2-oxoglutarate / glutamate / ATP / allosteric binding site
• Function / homology: 2-OXOGLUTARIC ACID / DI(HYDROXYETHYL)ETHER
• Biological species: Methanothermococcus thermolithotrophicus DSM 2095 (archaea)
• Method: X-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.91 Å
• Authors: Mueller, M.-C. / Wagner, T.

Funding support

Germany, 2items

Organization	Grant number	Country
Max Planck Society	na	Germany
German Research Foundation (DFG)	KU 3768/1-1	Germany

• Citation: Journal: Commun Biol / Year: 2024; Title: Differences in regulation mechanisms of glutamine synthetases from methanogenic archaea unveiled by structural investigations.; Authors: Muller, M.C. / Lemaire, O.N. / Kurth, J.M. / Welte, C.U. / Wagner, T.

History

Deposition	Apr 5, 2023	Deposition site: PDBE / Processing site: PDBE
Revision 1.0	Jan 24, 2024	Provider: repository / Type: Initial release
Revision 1.1	Jan 31, 2024	Group: Database references / Category: citation / citation_author Item: _citation.country / _citation.journal_abbrev / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.journal_volume / _citation.page_first / _citation.page_last / _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title / _citation.year / _citation_author.identifier_ORCID / _citation_author.name

Assembly

Deposited unit

A: Glutamine synthetase from Methanothermococcus thermolithotrophicus

B: Glutamine synthetase from Methanothermococcus thermolithotrophicus

C: Glutamine synthetase from Methanothermococcus thermolithotrophicus

D: Glutamine synthetase from Methanothermococcus thermolithotrophicus

E: Glutamine synthetase from Methanothermococcus thermolithotrophicus

F: Glutamine synthetase from Methanothermococcus thermolithotrophicus

G: Glutamine synthetase from Methanothermococcus thermolithotrophicus

H: Glutamine synthetase from Methanothermococcus thermolithotrophicus

I: Glutamine synthetase from Methanothermococcus thermolithotrophicus

J: Glutamine synthetase from Methanothermococcus thermolithotrophicus

K: Glutamine synthetase from Methanothermococcus thermolithotrophicus

L: Glutamine synthetase from Methanothermococcus thermolithotrophicus

P: Glutamine synthetase from Methanothermococcus thermolithotrophicus

Q: Glutamine synthetase from Methanothermococcus thermolithotrophicus

S: Glutamine synthetase from Methanothermococcus thermolithotrophicus

T: Glutamine synthetase from Methanothermococcus thermolithotrophicus

U: Glutamine synthetase from Methanothermococcus thermolithotrophicus

V: Glutamine synthetase from Methanothermococcus thermolithotrophicus

X: Glutamine synthetase from Methanothermococcus thermolithotrophicus

Y: Glutamine synthetase from Methanothermococcus thermolithotrophicus

Z: Glutamine synthetase from Methanothermococcus thermolithotrophicus

a: Glutamine synthetase from Methanothermococcus thermolithotrophicus

b: Glutamine synthetase from Methanothermococcus thermolithotrophicus

c: Glutamine synthetase from Methanothermococcus thermolithotrophicus

hetero molecules

Summary:

• 1.21 MDa, 24 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	1,212,788	92
Polymers	1,207,105	24
Non-polymers	5,683	68
Water	0	0

1

A: Glutamine synthetase from Methanothermococcus thermolithotrophicus

B: Glutamine synthetase from Methanothermococcus thermolithotrophicus

C: Glutamine synthetase from Methanothermococcus thermolithotrophicus

D: Glutamine synthetase from Methanothermococcus thermolithotrophicus

E: Glutamine synthetase from Methanothermococcus thermolithotrophicus

F: Glutamine synthetase from Methanothermococcus thermolithotrophicus

G: Glutamine synthetase from Methanothermococcus thermolithotrophicus

H: Glutamine synthetase from Methanothermococcus thermolithotrophicus

I: Glutamine synthetase from Methanothermococcus thermolithotrophicus

J: Glutamine synthetase from Methanothermococcus thermolithotrophicus

K: Glutamine synthetase from Methanothermococcus thermolithotrophicus

L: Glutamine synthetase from Methanothermococcus thermolithotrophicus

hetero molecules

Summary:

• defined by author&software
• Evidence: native gel electrophoresis, gel filtration
• 606 kDa, 12 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	605,786	36
Polymers	603,552	12
Non-polymers	2,234	24
Water	0

Symmetry operations:

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

Calculated values:

Buried area	63080 Å2
ΔGint	-283 kcal/mol
Surface area	194190 Å2
Method	PISA

2

P: Glutamine synthetase from Methanothermococcus thermolithotrophicus

Q: Glutamine synthetase from Methanothermococcus thermolithotrophicus

S: Glutamine synthetase from Methanothermococcus thermolithotrophicus

T: Glutamine synthetase from Methanothermococcus thermolithotrophicus

U: Glutamine synthetase from Methanothermococcus thermolithotrophicus

V: Glutamine synthetase from Methanothermococcus thermolithotrophicus

X: Glutamine synthetase from Methanothermococcus thermolithotrophicus

Y: Glutamine synthetase from Methanothermococcus thermolithotrophicus

Z: Glutamine synthetase from Methanothermococcus thermolithotrophicus

a: Glutamine synthetase from Methanothermococcus thermolithotrophicus

b: Glutamine synthetase from Methanothermococcus thermolithotrophicus

c: Glutamine synthetase from Methanothermococcus thermolithotrophicus

hetero molecules

Summary:

• defined by author&software
• 607 kDa, 12 polymers

Component details:

	Theoretical mass	Number of molelcules
Total (without water)	607,002	56
Polymers	603,552	12
Non-polymers	3,450	44
Water	0

Symmetry operations:

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

Calculated values:

Buried area	67710 Å2
ΔGint	-258 kcal/mol
Surface area	193150 Å2
Method	PISA

Unit cell

Length a, b, c (Å)	131.808, 131.927, 203.544
Angle α, β, γ (deg.)	89.95, 89.86, 60.05
Int Tables number	1
Space group name H-M	P1

Components

#1: Protein

A B C D E F G H I J K L P Q S T U V X ...
Glutamine synthetase from Methanothermococcus thermolithotrophicus

Details:

Mass: 50296.039 Da / Num. of mol.: 24 / Source method: isolated from a natural source
Source: (natural) Methanothermococcus thermolithotrophicus DSM 2095 (archaea)
Cell line: / / Organ: / / Plasmid details: / / Variant: / / Strain: DSM 2095 / Tissue: / / References: glutamine synthetase

#2: Chemical

A-501 E-501 F-501 G-501 K-501 P-501 P-503 S-601 U-501 U-502 V-501 V-502 X-601 X-602 Y-601 Y-602 Z-501 a-501 a-502 a-503 ...
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL

Details:

Mass: 62.068 Da / Num. of mol.: 25 / Source method: obtained synthetically / Formula: C₂H₆O₂

#3: Chemical

A-502 B-501 B-502 F-502 H-501 J-501 K-502 P-504 Q-501 Q-502 S-602 T-501 T-502 T-503 Y-603 b-503 b-504
ChemComp-MG / MAGNESIUM ION

Details:

Mass: 24.305 Da / Num. of mol.: 17 / Source method: obtained synthetically / Formula: Mg

#4: Chemical

A-503 B-503 C-501 D-501 E-502 F-503 G-502 H-502 I-501 J-502 K-503 L-501 P-505 Q-503 S-603 T-504 U-503 V-503 X-603 Y-604 ...
ChemComp-AKG / 2-OXOGLUTARIC ACID

Details:

Mass: 146.098 Da / Num. of mol.: 24 / Source method: obtained synthetically / Formula: C₅H₆O₅ / Feature type: SUBJECT OF INVESTIGATION

#5: Chemical

P-502 Z-502 ChemComp-PEG / DI(HYDROXYETHYL)ETHER

Details:

Mass: 106.120 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C₄H₁₀O₃

• Has ligand of interest: Y

Experimental details

Experiment

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

Sample preparation

• Crystal: Density Matthews: 2.54 ų/Da / Density % sol: 51.68 % / Description: Hexagonal plates
• Crystal grow: Temperature: 293.15 K / Method: vapor diffusion, sitting drop / pH: 5 ; Details: The protein was crystallized fresh without any freezing step, and crystallization was performed through the sitting drop method on a 96-Well MRC 2-Drop Crystallization Plates in polystyrene (SWISSCI, United Kingdom) under anaerobic conditions (N2:H2, gas ratio of 97:3). The enzyme was crystallized at 3.4 mg/ml with a final concentration of 2 mM 2-oxoglutarate and 2 mM MgCl in 25 mM Tris/HCl pH 7.6, 10% v/v glycerol, 150 mM NaCl, and 2 mM dithiothreitol. The crystallization reservoir contained 90 ul of mother liquor (25 % w/v Polyethylene glycol 1,500 and 100 mM SPG (succinic acid, sodium dihydrogen phosphate, and glycine) buffer pH 5.0), and the crystallization drop contained 0.6 ul protein with ligands and 0.6 ul precipitant. Crystals were soaked in the mother liquor supplemented with 15 % v/v glycerol prior to freezing in liquid nitrogen.

Data collection

• Diffraction: Mean temperature: 100 K / Serial crystal experiment: N
• Diffraction source: Source: SYNCHROTRON / Site: SLS / Beamline: X06DA / Wavelength: 0.99999 Å
• Detector: Type: DECTRIS PILATUS 2M-F / Detector: PIXEL / Date: Dec 10, 2020
• Radiation: Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
• Radiation wavelength: Wavelength: 0.99999 Å / Relative weight: 1
• Reflection: Resolution: 2.908→203.543 Å / Num. obs: 169224 / % possible obs: 91.7 % / Redundancy: 3.5 % / B_iso Wilson estimate: 51.2 Ų / CC_1/2: 0.993 / R_merge(I) obs: 0.122 / R_pim(I) all: 0.076 / R_rim(I) all: 0.144 / Net I/σ(I): 5.9
• Reflection shell: Resolution: 2.908→3.124 Å / Redundancy: 3 % / R_merge(I) obs: 0.501 / Mean I/σ(I) obs: 1.6 / Num. unique obs: 8461 / CC_1/2: 0.548 / R_pim(I) all: 0.344 / R_rim(I) all: 0.611 / % possible all: 69.3

Processing

Software

Name	Version	Classification
PHENIX	(1.20.1_4487: ???)	refinement
autoPROC		data reduction
STARANISO		data scaling
PHASER		phasing

Refinement

Method to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.91→43.74 Å / SU ML: 0.43 / Cross valid method: FREE R-VALUE / σ(F): 1.97 / Phase error: 30.68 / Stereochemistry target values: ML
Details: Refinement was performed with PHENIX and BUSTER in combination with fast automatic visual model building in COOT. The model was systematically validated by using Molprobity. The model was refined in PHENIX with translation libration screw model and without non-crystallography symmetry and with riding hydrogens. Hydrogens were omitted in the deposited model.

	Rfactor	Num. reflection	% reflection
Rfree	0.278	8324	4.92 %
Rwork	0.2541	-	-
obs	0.2553	169113	64.78 %

• Solvent computation: Shrinkage radii: 0.9 Å / VDW probe radii: 1.1 Å / Solvent model: FLAT BULK SOLVENT MODEL

Refinement step

Cycle: LAST / Resolution: 2.91→43.74 Å

	Protein	Nucleic acid	Ligand	Solvent	Total
Num. atoms	84792	0	371	0	85163

Refine LS restraints

Refine-ID	Type	Dev ideal	Number
X-RAY DIFFRACTION	f_bond_d	0.01	87279
X-RAY DIFFRACTION	f_angle_d	1.274	118180
X-RAY DIFFRACTION	f_dihedral_angle_d	13.228	32423
X-RAY DIFFRACTION	f_chiral_restr	0.081	12620
X-RAY DIFFRACTION	f_plane_restr	0.005	15596

LS refinement shell

Resolution (Å)	Rfactor Rfree	Num. reflection Rfree	Rfactor Rwork	Num. reflection Rwork	Refine-ID	% reflection obs (%)
2.91-2.94	0.2683	32	0.3551	567	X-RAY DIFFRACTION	7
2.94-2.98	0.4271	43	0.3588	968	X-RAY DIFFRACTION	12
2.98-3.01	0.4152	60	0.3523	1412	X-RAY DIFFRACTION	17
3.01-3.05	0.3492	97	0.3224	1577	X-RAY DIFFRACTION	19
3.05-3.09	0.4034	71	0.3153	1880	X-RAY DIFFRACTION	22
3.09-3.13	0.3798	137	0.3196	2138	X-RAY DIFFRACTION	26
3.13-3.18	0.346	111	0.3344	2426	X-RAY DIFFRACTION	29
3.18-3.23	0.337	170	0.3153	2827	X-RAY DIFFRACTION	34
3.23-3.28	0.3813	228	0.3105	2984	X-RAY DIFFRACTION	37
3.28-3.33	0.3573	170	0.3103	3384	X-RAY DIFFRACTION	41
3.33-3.39	0.3082	240	0.3172	3766	X-RAY DIFFRACTION	46
3.39-3.45	0.3075	231	0.2977	4031	X-RAY DIFFRACTION	49
3.45-3.51	0.261	201	0.2871	4358	X-RAY DIFFRACTION	52
3.51-3.59	0.294	305	0.2837	4714	X-RAY DIFFRACTION	58
3.59-3.66	0.3215	312	0.2828	5504	X-RAY DIFFRACTION	66
3.66-3.75	0.3372	293	0.285	6232	X-RAY DIFFRACTION	75
3.75-3.84	0.3067	332	0.2737	6810	X-RAY DIFFRACTION	83
3.84-3.95	0.3173	439	0.2815	7334	X-RAY DIFFRACTION	90
3.95-4.06	0.2931	404	0.2763	7971	X-RAY DIFFRACTION	96
4.06-4.19	0.2688	362	0.2525	8192	X-RAY DIFFRACTION	98
4.19-4.34	0.2806	325	0.2408	8272	X-RAY DIFFRACTION	99
4.34-4.52	0.2762	400	0.2286	8193	X-RAY DIFFRACTION	99
4.52-4.72	0.2411	301	0.2235	8360	X-RAY DIFFRACTION	99
4.72-4.97	0.2362	366	0.2276	8192	X-RAY DIFFRACTION	98
4.97-5.28	0.2657	535	0.2336	8151	X-RAY DIFFRACTION	99
5.28-5.69	0.2629	288	0.2467	8286	X-RAY DIFFRACTION	99
5.69-6.26	0.3116	447	0.256	8114	X-RAY DIFFRACTION	99
6.26-7.16	0.2741	473	0.2481	8104	X-RAY DIFFRACTION	99
7.16-9.01	0.246	404	0.2282	8130	X-RAY DIFFRACTION	98
9.01-43.74	0.2044	547	0.2196	7912	X-RAY DIFFRACTION	97

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	0.365	0.142	-0.0174	0.8182	-0.4997	0.5916	0.0233	0.4707	0.4334	-0.2495	-0.0422	-0.1213	-0.1894	0.0214	-0.0116	1.0186	0.4471	0.1758	1.1284	0.3956	-0.0629	31.1436	22.2833	-31.0159
2	0.7029	0.486	-0.6632	0.4036	-0.2104	1.1836	-0.1124	-0.2778	-0.0749	0.0574	-0.2166	-0.3876	0.4441	0.384	0.3355	0.8636	0.2428	0.0716	0.983	0.245	0.478	47.9666	22.1463	-15.638
3	0.8781	-0.1338	-0.4712	0.454	0.4459	1.5725	0.015	-0.2053	-0.0745	0.1895	-0.0475	0.1699	0.1755	-0.1127	0.0163	0.8391	0.3306	0.2318	0.9287	0.1131	0.2773	-34.2935	-16.775	30.9366
4	0.8423	0.5715	-0.8566	0.3964	-0.7382	2.9218	-0.2577	-0.0842	-0.4978	-0.0817	0.0204	-0.0711	1.2683	-0.1265	0.246	1.1659	0.3009	0.13	0.9363	0.0029	0.3615	-43.0695	-30.7924	15.2511
5	1.2656	0.6036	0.3762	1.1395	0.0684	0.8098	-0.0607	0.4187	0.1265	-0.3749	0.0653	0.3048	-0.058	-0.1669	-0.0014	0.8768	0.4054	-0.0459	1.1233	0.1208	0.3122	-35.2263	16.8648	-30.0543
6	1.7048	-0.3139	0.6015	0.5341	-0.7916	1.5935	-0.0802	-0.0198	0.6411	0.0648	0.055	0.033	-0.8016	-0.2206	0.0701	1.093	0.5742	0.0759	0.9101	0.0164	0.5542	-43.2753	30.9202	-13.9973
7	1.043	0.2532	0.015	1.0053	0.2361	1.3596	0.1609	-0.4653	0.1429	0.3445	-0.1563	-0.1526	-0.3507	0.154	-0.1008	1.0818	0.1566	-0.066	0.9521	-0.1833	0.3769	19.7923	35.7451	36.535
8	1.1892	0.3582	0.1739	0.8426	0.2237	0.7317	0.0469	-0.2706	0.2942	0.1907	-0.0454	0.0512	-0.39	-0.0743	-0.0506	0.9556	0.1854	0.0505	0.9013	-0.229	0.416	0.6374	35.9092	29.495
9	0.5561	0.2864	-0.1234	0.9054	0.5619	0.5544	0.1696	-0.3708	0.5847	0.1873	-0.206	0.2427	-0.5648	-0.2663	0.0216	1.318	0.4053	0.0482	1.009	-0.2439	0.704	-8.6152	53.8386	26.2704
10	0.6605	-0.4184	-0.9121	2.1041	1.0811	1.5157	0.1564	0.0321	0.6161	-0.1213	-0.3131	0.5267	-0.3874	-0.664	0.1245	1.1548	0.4178	0.0083	0.7784	-0.1493	0.7901	-4.1576	51.6228	14.85
11	1.0576	-0.0029	0.2565	0.4618	-0.297	1.716	0.0382	-0.3617	0.0397	0.296	-0.2311	-0.2748	-0.3262	0.2868	0.0919	1.0759	0.1508	-0.1557	0.9527	-0.0501	0.0224	36.5734	17.1074	29.5998
12	1.0829	1.6163	0.4463	3.5756	1.1041	0.3499	-0.1464	0.2611	0.5652	-0.0564	-0.2139	0.1116	-1.0416	0.2499	0.3854	1.367	-0.0041	-0.1479	1.0286	0.0855	0.4326	37.0876	33.7519	13.139
13	0.4509	0.2663	0.8824	0.1469	0.4914	2.063	-0.0769	-0.1529	0.3874	0.2835	-0.1048	-0.0724	-0.6468	-0.0655	0.1863	1.0885	0.1258	-0.0831	1.0001	0.0134	0.3585	44.2462	28.6737	13.621
14	1.1389	0.3159	0.0799	1.0538	0.4611	1.6162	0.1214	-0.3135	0.4358	0.2082	-0.0387	0.1098	0.0227	-0.1835	-0.0974	0.8758	0.3548	0.1648	0.7578	-0.0699	0.3612	-32.486	21.7663	30.8882
15	0.4253	-0.3668	-0.3703	0.921	-0.3211	1.2606	0.0949	0.2744	-0.2115	0.0424	-0.1934	0.3603	0.0107	-0.7702	0.0361	0.8735	0.4214	0.0543	0.8686	0.0242	0.4574	-46.81	21.6779	15.123
16	0.3924	-0.0563	0.0221	0.6775	-0.1599	0.6376	-0.0027	-0.2042	-0.3604	0.1962	-0.0534	-0.0257	0.1093	0.1111	0.0625	0.7983	0.3822	0.0196	1.1046	0.3144	0.1748	29.2257	-23.7972	30.3002
17	0.2826	-0.2072	0.1261	0.3936	-0.3119	0.5216	0.0129	-0.2695	-0.0493	0.3593	-0.1386	-0.2404	-0.0739	0.3626	0.1739	0.6779	0.2202	-0.0271	1.164	0.3726	0.3586	52.0677	-19.723	24.0234
18	2.6649	0.3506	1.8101	0.0987	-0.0358	2.6822	-0.2161	-0.0901	0.1999	0.2133	-0.1666	-0.2086	-0.9816	0.4083	0.3764	0.8786	0.272	0.1094	1.0184	0.2687	0.4888	48.0983	-15.1622	12.3164
19	0.276	-0.0969	0.5873	0.2538	-0.6079	2.163	-0.0456	-0.1337	-0.0097	0.0135	-0.2267	-0.302	-0.3299	0.6422	0.2895	0.7979	0.3797	0.0964	0.9552	0.3189	0.2725	47.2737	-23.9053	12.8666
20	1.0009	0.3064	-0.6382	0.549	0.0676	1.005	-0.0784	-0.2141	-0.2518	0.1421	-0.0296	0.1454	0.1147	0.0173	0.1076	0.9017	0.4335	0.1736	0.8848	0.1927	0.1855	-5.1477	-36.9327	30.665
21	0.182	-0.0749	0.3323	1.0188	-0.8563	1.1604	-0.024	-0.1093	-0.2569	-0.1787	-0.0893	-0.22	0.4806	0.6179	0.1391	1.0866	0.4634	0.1855	0.8581	0.1693	0.4654	6.077	-53.0132	16.2549
22	0.3207	0.3294	0.096	1.1413	0.4012	1.5444	0.0843	0.2974	-0.2393	-0.3638	0.0002	0.0763	0.1143	-0.3269	-0.0872	0.8501	0.2956	-0.1082	1.2227	-0.2577	0.2688	-33.9275	-21.7967	-29.8505
23	0.5633	0.276	1.0065	0.1725	0.1881	2.906	-0.1898	0.1158	0.1299	-0.0498	-0.2295	0.4715	-0.2401	-1.0517	0.3925	0.8464	0.3573	-0.1023	1.34	-0.2653	0.5821	-47.6644	-21.4713	-13.6002
24	1.8116	0.2362	0.3721	1.4845	0.2853	1.2463	-0.1024	0.4411	0.5996	-0.3064	0.0615	0.1878	-0.1684	0.0735	-0.0217	0.8817	0.2923	0.0717	0.9056	0.3228	0.4954	-2.843	40.4055	-29.5803
25	3.3385	0.9299	-1.2589	6.1298	-1.483	0.6921	-0.1314	-0.2449	-0.0239	0.4716	-0.4743	-0.3371	-0.7798	0.9679	0.6125	1.0404	0.1913	0.0153	0.9509	0.2711	0.6923	11.3085	49.0692	-13.1404
26	0.9873	0.3895	-0.1423	1.3907	-0.4644	2.1038	-0.2486	0.2378	0.4673	0.2662	0.182	0.029	-0.4662	0.0438	0.0574	1.0609	0.2835	0.0304	0.7021	0.1754	0.7142	3.3347	52.759	-13.6388
27	0.7798	-0.1556	-0.3288	0.5497	0.0123	0.7587	-0.1246	0.3358	-0.3295	-0.3101	0.0081	-0.0885	0.2926	-0.16	0.078	1.1292	0.2944	0.0373	0.9304	-0.3119	0.1937	1.7862	-39.7565	-30.6527
28	0.567	0.2477	0.144	1.367	1.1482	2.4176	0.1286	0.14	-0.1918	-0.1923	-0.1705	0.269	0.1064	-0.5641	0.0536	1.0337	0.3742	0.0153	0.8384	-0.0355	0.5	-5.2946	-51.8209	-14.633
29	0.5901	0.2271	-0.1612	0.4561	-0.0945	0.8333	-0.0704	0.3488	-0.0861	-0.281	-0.0564	-0.4115	0.191	0.0432	0.0877	1.0894	0.4199	0.279	1.1335	0.0024	0.0582	35.0257	-17.4881	-31.0079
30	1.7465	-0.1282	-1.7534	0.5465	0.5902	3.0399	-0.3009	0.2029	-0.5535	0.0624	-0.0245	-0.0364	1.1595	0.085	0.3873	1.0697	0.3951	0.2207	0.958	0.1838	0.4239	41.9853	-30.0187	-15.245
31	1.0693	0.4032	0.4152	0.9275	0.1261	0.4754	-0.12	-0.3504	0.3198	0.2864	0.0145	-0.1623	-0.1124	-0.1062	0.0829	0.3554	-0.0282	-0.1854	0.3307	-0.2171	0.6024	12.4149	-10.9024	132.4394
32	0.3474	0.2004	0.0099	0.917	0.4388	0.8279	-0.1904	-0.271	0.307	0.2296	0.1849	0.2957	-0.7043	-0.5072	0.1704	0.1988	-0.2004	-0.1944	0.1156	-0.2699	0.9473	1.0587	5.1608	118.1002
33	0.7169	0.2345	-0.1213	1.8628	-0.3432	0.8703	-0.0085	0.4576	-0.4851	-0.3201	0.0631	-0.304	0.2797	-0.0888	-0.092	0.3438	-0.0894	0.0319	0.3642	-0.2906	0.8559	9.9024	-88.0021	72.1847
34	0.2664	0.3144	0.6087	1.6068	1.1709	1.4039	-0.1142	0.1367	-0.2884	0.0148	-0.0497	0.5033	0.7645	-0.764	0.1338	0.4899	-0.4704	0.078	0.1574	-0.2496	1.2351	2.3644	-99.7484	88.2261
35	0.9403	-0.5945	-0.3086	1.5777	-0.1202	0.6543	-0.0816	-0.5814	-0.2907	0.4463	0.1286	-0.1547	0.2333	0.1265	-0.0197	0.5123	0.0204	-0.2615	0.3189	0.1672	0.7988	35.2917	-70.3113	133.6268
36	0.6362	-0.0038	-0.0296	0.061	0.1843	2.188	-0.0133	-0.2457	0.2102	0.2031	0.0136	-0.4677	-0.3651	1.0552	-0.0407	0.2869	-0.1067	-0.2354	0.3181	0.0166	1.2386	55.3049	-68.5662	119.8352
37	0.9264	0.0726	0.0949	1.1925	-0.1066	1.3128	0.0301	0.4188	0.2197	-0.3883	-0.0129	-0.2546	-0.1762	0.2651	-0.049	0.3235	-0.154	0.1164	0.5342	0.1721	0.5543	39.2539	-26.1159	71.0894
38	5.18	0.8063	-2.2153	1.4745	0.3402	1.7576	0.109	-0.1777	-0.6802	0.0907	-0.1747	-0.7171	0.1314	1.1186	0.1712	0.0809	-0.2701	0.1074	0.8092	0.3118	1.0846	59.9618	-29.2328	83.1692
39	0.5027	0.1152	-0.3891	0.771	-0.4254	0.8617	-0.053	0.2924	0.2126	-0.0896	0.0941	-0.1617	-0.3034	0.1737	-0.1018	0.0386	-0.3559	0.0129	0.1157	0.1527	0.7083	46.1155	-18.6316	95.2795
40	1.2954	-0.0523	0.4051	1.5318	0.0285	1.8164	-0.04	0.3864	0.1805	-0.5342	-0.0814	0.1681	-0.1415	-0.2024	0.0848	0.6743	-0.0475	-0.1511	0.3137	0.2276	0.5805	-11.0674	-11.6624	64.6237
41	0.8997	0.3212	0.1373	0.9393	0.1253	0.8826	0.0053	0.2015	0.363	-0.4589	-0.0208	0.0165	-0.279	-0.0246	0.0578	0.6672	-0.0587	-0.1013	0.3615	0.2332	0.5416	-4.23	-3.5101	66.0803
42	1.1702	-0.5368	0.3373	0.8869	0.3055	0.4846	-0.098	0.2163	0.2999	-0.3998	0.0105	-0.1789	-0.1271	0.0942	0.0319	0.4379	-0.0648	-0.0215	0.2692	0.1538	0.5754	14.1638	-15.7167	75.3935
43	0.0604	-0.039	-0.1862	1.6859	-0.7455	1.5608	0.061	0.2889	0.4176	-0.3626	-0.028	-0.4266	-0.6681	0.6106	0.0345	0.6476	-0.1716	-0.0595	0.2918	0.1775	1.0532	16.3984	5.0665	78.1635
44	0.0177	0.1168	-0.1836	1.3565	-0.4711	1.6936	-0.012	0.112	0.3361	-0.2219	0.0247	-0.3952	-0.6138	0.4003	0.0182	0.3824	-0.2382	-0.0775	0.195	0.0549	1.113	9.3453	4.1508	92.9025
45	1.2224	0.1301	0.3557	0.8941	-0.6362	0.8686	0.0466	0.4619	-0.3666	-0.4302	-0.1187	-0.4276	0.1793	0.2902	0.0698	0.3441	-0.0123	0.1631	0.3471	-0.1462	0.8052	43.2129	-65.8905	72.2512
46	0.6461	-0.5917	-0.3496	0.6571	-0.126	1.9284	0.1	0.2149	-0.5033	-0.2526	-0.2476	-0.0977	0.7236	0.2169	0.1308	0.2012	0.2313	-0.0632	0.2383	-0.1372	1.195	49.6464	-78.0171	88.5035
47	2.0431	0.0205	0.0755	1.1614	0.337	1.5429	-0.0095	0.5682	0.2084	-0.6289	-0.0827	0.1511	-0.2023	-0.1672	0.0802	0.4978	-0.0729	-0.2009	0.4138	0.0066	0.3982	-27.0841	-31.5802	70.2628
48	0.6164	0.4211	0.1875	0.8024	0.3821	2.7359	-0.0434	0.2298	0.7662	-0.3399	-0.147	0.399	-0.8263	-0.1364	0.236	0.4035	-0.0403	-0.1902	0.3754	-0.0741	0.9421	-35.6858	-17.1563	85.7191
49	1.014	0.1972	0.1098	1.2891	-0.021	0.7576	0.0851	0.5975	-0.5404	-0.4206	-0.0719	0.1055	0.2397	-0.0476	-0.0594	0.3207	-0.0816	-0.1101	0.4692	-0.2513	0.4982	-21.2738	-71.0391	70.3331
50	1.8722	0.9765	1.1041	0.6631	0.2416	1.1599	0.1925	0.0896	0.1473	-0.2387	-0.2465	0.4684	-0.4428	-0.8411	0.2594	0.0651	-0.3528	-0.1783	0.1795	-0.3398	0.7824	-41.2152	-68.7799	84.1338
51	0.7168	0.2525	-0.3115	0.8433	-0.1001	0.5936	-0.0027	-0.449	-0.3793	0.617	0.0845	-0.0812	0.4218	-0.1322	-0.0428	0.6514	-0.2707	-0.0649	0.3348	0.3503	0.7416	4.0041	-87.1291	132.3104
52	0.2186	0.4756	-0.0437	1.7143	-0.5272	0.8028	0.0909	-0.0993	-0.4154	0.385	0.0104	-0.8053	0.355	0.4299	0.0268	0.3268	-0.1251	-0.0971	0.2256	0.2301	1.2238	11.5511	-99.3107	116.5372
53	0.9267	-0.2487	0.0539	0.415	-0.401	0.604	-0.1194	-0.3933	0.2064	0.5609	-0.008	-0.2516	-0.1485	0.1452	0.0586	0.0674	-0.1545	-0.4131	0.2133	-0.1248	0.773	41.6158	-30.9267	132.5633
54	0.4997	0.4736	0.6756	0.6633	0.6969	1.2427	-0.0661	-0.0452	0.5262	0.189	-0.0894	-0.1304	-0.6678	0.1583	0.1362	0.2071	-0.2827	-0.2418	0.3443	-0.0143	1.058	50.2698	-16.9816	116.8614
55	1.0433	-0.2993	-0.0482	0.9151	0.1315	1.2174	-0.1468	-0.5955	0.0957	0.5648	0.0071	0.2527	-0.1138	-0.2428	0.0719	0.7203	-0.3182	0.3247	0.7815	-0.1522	0.2519	-32.5499	-50.397	137.9883
56	0.9267	-0.2062	0.4394	0.5576	0.2565	0.4947	-0.0251	-0.2846	-0.3245	0.4873	-0.0343	0.2215	0.2138	-0.1454	0.0422	0.6558	-0.2333	0.1917	0.4195	0.0822	0.529	-25.8196	-70.9013	128.9251
57	0.81	0.5939	-0.4576	0.7817	-0.8321	0.8748	-0.0558	-0.0352	-0.7082	0.1709	0.027	0.198	0.7067	-0.0679	0.1058	0.3062	-0.747	0.3285	0.2742	0.0339	0.6325	-36.3711	-77.9892	116.4612
58	1.1276	-0.1594	0.0758	1.1936	-0.1892	0.9031	-0.067	-0.4234	0.3205	0.4123	-0.0753	0.1964	-0.0179	-0.2642	0.1224	0.2707	-0.058	0.0484	0.5046	-0.2732	0.4467	-26.4599	-24.847	130.9618
59	0.9706	-0.2904	-0.8023	0.1551	-0.1554	2.0156	0.0435	-0.1712	-0.1316	0.0961	-0.1809	0.6625	0.3471	-1.04	0.1677	0.1727	0.03	-0.0178	0.4937	-0.2601	0.9592	-40.2362	-25.4981	114.7353

Refinement TLS group

ID	Refine-ID	Refine TLS-ID	Selection details
1	X-RAY DIFFRACTION	1	chain 'A' and (resid 3 through 272 )
2	X-RAY DIFFRACTION	2	chain 'A' and (resid 273 through 448 )
3	X-RAY DIFFRACTION	3	chain 'B' and (resid 3 through 272 )
4	X-RAY DIFFRACTION	4	chain 'B' and (resid 273 through 448 )
5	X-RAY DIFFRACTION	5	chain 'C' and (resid 2 through 272 )
6	X-RAY DIFFRACTION	6	chain 'C' and (resid 273 through 448 )
7	X-RAY DIFFRACTION	7	chain 'D' and (resid 2 through 73 )
8	X-RAY DIFFRACTION	8	chain 'D' and (resid 74 through 236 )
9	X-RAY DIFFRACTION	9	chain 'D' and (resid 237 through 295 )
10	X-RAY DIFFRACTION	10	chain 'D' and (resid 296 through 448 )
11	X-RAY DIFFRACTION	11	chain 'E' and (resid 2 through 295 )
12	X-RAY DIFFRACTION	12	chain 'E' and (resid 296 through 323 )
13	X-RAY DIFFRACTION	13	chain 'E' and (resid 324 through 448 )
14	X-RAY DIFFRACTION	14	chain 'F' and (resid 2 through 295 )
15	X-RAY DIFFRACTION	15	chain 'F' and (resid 296 through 448 )
16	X-RAY DIFFRACTION	16	chain 'G' and (resid 2 through 236 )
17	X-RAY DIFFRACTION	17	chain 'G' and (resid 237 through 295 )
18	X-RAY DIFFRACTION	18	chain 'G' and (resid 296 through 323 )
19	X-RAY DIFFRACTION	19	chain 'G' and (resid 324 through 448 )
20	X-RAY DIFFRACTION	20	chain 'H' and (resid 2 through 236 )
21	X-RAY DIFFRACTION	21	chain 'H' and (resid 237 through 448 )
22	X-RAY DIFFRACTION	22	chain 'I' and (resid 2 through 295 )
23	X-RAY DIFFRACTION	23	chain 'I' and (resid 296 through 448 )
24	X-RAY DIFFRACTION	24	chain 'J' and (resid 2 through 295 )
25	X-RAY DIFFRACTION	25	chain 'J' and (resid 296 through 323 )
26	X-RAY DIFFRACTION	26	chain 'J' and (resid 324 through 448 )
27	X-RAY DIFFRACTION	27	chain 'K' and (resid 2 through 295 )
28	X-RAY DIFFRACTION	28	chain 'K' and (resid 296 through 448 )
29	X-RAY DIFFRACTION	29	chain 'L' and (resid 2 through 295 )
30	X-RAY DIFFRACTION	30	chain 'L' and (resid 296 through 448 )
31	X-RAY DIFFRACTION	31	chain 'P' and (resid 2 through 236 )
32	X-RAY DIFFRACTION	32	chain 'P' and (resid 237 through 448 )
33	X-RAY DIFFRACTION	33	chain 'Q' and (resid 3 through 295 )
34	X-RAY DIFFRACTION	34	chain 'Q' and (resid 296 through 448 )
35	X-RAY DIFFRACTION	35	chain 'S' and (resid 2 through 236 )
36	X-RAY DIFFRACTION	36	chain 'S' and (resid 237 through 448 )
37	X-RAY DIFFRACTION	37	chain 'T' and (resid 2 through 272 )
38	X-RAY DIFFRACTION	38	chain 'T' and (resid 273 through 395 )
39	X-RAY DIFFRACTION	39	chain 'T' and (resid 396 through 448 )
40	X-RAY DIFFRACTION	40	chain 'U' and (resid 2 through 73 )
41	X-RAY DIFFRACTION	41	chain 'U' and (resid 74 through 128 )
42	X-RAY DIFFRACTION	42	chain 'U' and (resid 129 through 236 )
43	X-RAY DIFFRACTION	43	chain 'U' and (resid 237 through 366 )
44	X-RAY DIFFRACTION	44	chain 'U' and (resid 367 through 448 )
45	X-RAY DIFFRACTION	45	chain 'V' and (resid 2 through 295 )
46	X-RAY DIFFRACTION	46	chain 'V' and (resid 296 through 448 )
47	X-RAY DIFFRACTION	47	chain 'X' and (resid 2 through 272 )
48	X-RAY DIFFRACTION	48	chain 'X' and (resid 273 through 448 )
49	X-RAY DIFFRACTION	49	chain 'Y' and (resid 2 through 236 )
50	X-RAY DIFFRACTION	50	chain 'Y' and (resid 237 through 448 )
51	X-RAY DIFFRACTION	51	chain 'Z' and (resid 2 through 295 )
52	X-RAY DIFFRACTION	52	chain 'Z' and (resid 296 through 448 )
53	X-RAY DIFFRACTION	53	chain 'a' and (resid 3 through 272 )
54	X-RAY DIFFRACTION	54	chain 'a' and (resid 273 through 448 )
55	X-RAY DIFFRACTION	55	chain 'b' and (resid 2 through 99 )
56	X-RAY DIFFRACTION	56	chain 'b' and (resid 100 through 272 )
57	X-RAY DIFFRACTION	57	chain 'b' and (resid 273 through 448 )
58	X-RAY DIFFRACTION	58	chain 'c' and (resid 2 through 295 )
59	X-RAY DIFFRACTION	59	chain 'c' and (resid 296 through 448 )