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- PDB-4ohs: The structure of a far-red fluorescent protein, AQ143 -

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Basic information

Entry
Database: PDB / ID: 4ohs
TitleThe structure of a far-red fluorescent protein, AQ143
ComponentsFAR-RED FLUORESCENT PROTEIN AQ143
KeywordsFLUORESCENT PROTEIN / far-red / Beta Barrel / Red Fluorescent Protein / RFP
Function / homologyGreen Fluorescent Protein / Green fluorescent protein / Green fluorescent protein-related / Green fluorescent protein / Green fluorescent protein / bioluminescence / Beta Barrel / Mainly Beta / Blue chromoprotein aeCP597
Function and homology information
Biological speciesActinia Equina (sea anemone)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.19 Å
AuthorsWannier, T.M. / Mayo, S.L.
CitationJournal: Protein Sci. / Year: 2014
Title: The structure of a far-red fluorescent protein, AQ143, shows evidence in support of reported red-shifting chromophore interactions.
Authors: Wannier, T.M. / Mayo, S.L.
History
DepositionJan 17, 2014Deposition site: RCSB / Processing site: RCSB
Revision 1.0Feb 26, 2014Provider: repository / Type: Initial release
Revision 1.1Jul 2, 2014Group: Database references
Revision 1.2Aug 6, 2014Group: Database references

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: FAR-RED FLUORESCENT PROTEIN AQ143
B: FAR-RED FLUORESCENT PROTEIN AQ143
C: FAR-RED FLUORESCENT PROTEIN AQ143
D: FAR-RED FLUORESCENT PROTEIN AQ143
E: FAR-RED FLUORESCENT PROTEIN AQ143
F: FAR-RED FLUORESCENT PROTEIN AQ143
G: FAR-RED FLUORESCENT PROTEIN AQ143
H: FAR-RED FLUORESCENT PROTEIN AQ143
hetero molecules


Theoretical massNumber of molelcules
Total (without water)213,66412
Polymers213,5228
Non-polymers1424
Water6,395355
1
A: FAR-RED FLUORESCENT PROTEIN AQ143
B: FAR-RED FLUORESCENT PROTEIN AQ143
D: FAR-RED FLUORESCENT PROTEIN AQ143
E: FAR-RED FLUORESCENT PROTEIN AQ143
hetero molecules


Theoretical massNumber of molelcules
Total (without water)106,8677
Polymers106,7614
Non-polymers1063
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area11430 Å2
ΔGint-83 kcal/mol
Surface area31340 Å2
MethodPISA
2
C: FAR-RED FLUORESCENT PROTEIN AQ143
F: FAR-RED FLUORESCENT PROTEIN AQ143
G: FAR-RED FLUORESCENT PROTEIN AQ143
H: FAR-RED FLUORESCENT PROTEIN AQ143
hetero molecules


Theoretical massNumber of molelcules
Total (without water)106,7975
Polymers106,7614
Non-polymers351
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area11270 Å2
ΔGint-65 kcal/mol
Surface area31050 Å2
MethodPISA
Unit cell
Length a, b, c (Å)50.967, 68.114, 132.788
Angle α, β, γ (deg.)98.38, 90.74, 110.47
Int Tables number1
Space group name H-MP1

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Components

#1: Protein
FAR-RED FLUORESCENT PROTEIN AQ143


Mass: 26690.262 Da / Num. of mol.: 8 / Source method: isolated from a natural source / Source: (natural) Actinia Equina (sea anemone) / References: UniProt: Q2VFP3*PLUS
#2: Chemical
ChemComp-CL / CHLORIDE ION / Chloride


Mass: 35.453 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Cl
#3: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 355 / Source method: isolated from a natural source / Formula: H2O

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION

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Sample preparation

CrystalDensity Matthews: 2 Å3/Da / Density % sol: 38.39 %
Crystal growTemperature: 298 K / Method: vapor diffusion, hanging drop / pH: 7
Details: 100mM Tris, 50mM lithium sulfate, 20% w/v PEG 3350, pH 7.0, VAPOR DIFFUSION, HANGING DROP, temperature 298K

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Data collection

Diffraction sourceSource: SYNCHROTRON / Site: SSRL / Beamline: BL12-2 / Wavelength: 1
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 2.19→131.1 Å / Num. obs: 72946

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Processing

SoftwareName: PHENIX / Version: (phenix.refine: 1.8.4_1496) / Classification: refinement
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.19→37.326 Å / SU ML: 0.26 / σ(F): 1.97 / Phase error: 30.84 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2205 3647 5 %
Rwork0.1897 --
obs0.1912 72881 86.08 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.19→37.326 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms14128 0 4 355 14487
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00614596
X-RAY DIFFRACTIONf_angle_d0.98719737
X-RAY DIFFRACTIONf_dihedral_angle_d15.6755398
X-RAY DIFFRACTIONf_chiral_restr0.0412031
X-RAY DIFFRACTIONf_plane_restr0.0062591
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.19-2.22130.3391600.31541023X-RAY DIFFRACTION33
2.2213-2.25170.34291040.29532013X-RAY DIFFRACTION65
2.2517-2.28390.35481290.28982334X-RAY DIFFRACTION76
2.2839-2.3180.33741320.27182814X-RAY DIFFRACTION90
2.318-2.35420.31361610.2692771X-RAY DIFFRACTION91
2.3542-2.39280.3191650.26572828X-RAY DIFFRACTION91
2.3928-2.43410.29761440.25692788X-RAY DIFFRACTION90
2.4341-2.47830.29061250.2492814X-RAY DIFFRACTION91
2.4783-2.5260.2961470.25112793X-RAY DIFFRACTION90
2.526-2.57750.30071400.24782755X-RAY DIFFRACTION88
2.5775-2.63350.28031220.24422662X-RAY DIFFRACTION87
2.6335-2.69480.28241240.2382686X-RAY DIFFRACTION85
2.6948-2.76220.2911460.21982836X-RAY DIFFRACTION92
2.7622-2.83680.2521590.22242831X-RAY DIFFRACTION91
2.8368-2.92030.26651470.2362795X-RAY DIFFRACTION92
2.9203-3.01450.2571610.23352825X-RAY DIFFRACTION91
3.0145-3.12220.25691430.22522706X-RAY DIFFRACTION88
3.1222-3.24710.24561200.21232624X-RAY DIFFRACTION84
3.2471-3.39480.25381490.19982836X-RAY DIFFRACTION91
3.3948-3.57360.22881630.17952814X-RAY DIFFRACTION92
3.5736-3.79730.20281650.17012805X-RAY DIFFRACTION91
3.7973-4.09020.18881610.16662757X-RAY DIFFRACTION90
4.0902-4.50120.16051460.13932674X-RAY DIFFRACTION87
4.5012-5.15110.15881590.12852893X-RAY DIFFRACTION93
5.1511-6.48430.16761400.14982683X-RAY DIFFRACTION87
6.4843-37.33170.16371350.16612874X-RAY DIFFRACTION92
Refinement TLS params.

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

IDL112)L122)L132)L222)L232)L332)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T112)T122)T132)T222)T232)T332)Origin x (Å)Origin y (Å)Origin z (Å)
16.00475.3407-2.26127.4179-3.24415.2103-0.40820.15570.0012-0.67170.32270.63210.7451-0.39830.03830.4852-0.07860.00540.3587-0.04220.335429.061323.7925.7943
20.5325-0.0468-0.50682.29450.00622.5264-0.0295-0.00480.01530.02430.08440.34120.4403-0.2279-0.05520.4215-0.09820.07520.3369-0.02090.320330.737125.627134.6249
39.6713-0.16165.43614.78130.26635.01740.0536-0.24640.10610.3126-0.11230.17350.2230.11340.08770.4093-0.02280.13580.2437-0.01730.244835.021628.832743.0604
42.1541-1.13230.55514.3077-0.35565.1348-0.0909-0.0978-0.41450.21010.07310.75670.4398-0.2837-0.02330.6956-0.21510.06060.3169-0.07980.498625.968219.942540.5158
58.60824.377-8.21348.0883-6.52222.0893-0.5073-0.4745-0.54070.1297-0.1243-0.43371.04570.53680.76220.4809-0.0354-0.02010.3881-0.01470.382335.484717.521435.589
61.52390.3083-0.1382.13370.51231.7166-0.08080.21920.1132-0.28090.05550.2867-0.3401-0.05750.01730.3753-0.05140.04380.32810.02040.377827.883353.149832.1375
79.4561.3793-5.74783.3053-1.07717.966-0.15260.1113-0.1463-0.24440.09860.0654-0.03680.1930.10490.3669-0.05780.0120.1976-0.02340.254236.580652.812638.7379
86.43872.49836.94476.53722.33539.8311-0.5662-0.77180.6999-0.09240.11530.5143-1.0677-0.60150.45750.5855-0.0210.18510.3552-0.01520.400429.666762.476239.6049
90.7365-0.51961.72421.0502-1.25442.09340.00470.35360.1239-0.01680.2467-0.2521-1.52191.1798-0.33991.0057-0.30190.27120.7821-0.11640.533950.416651.60242.8063
101.6097-0.36271.34163.2912-2.6175.90850.014-0.19860.3240.25890.3011-0.0089-2.0469-0.0205-0.30051.1665-0.1850.30810.5143-0.10050.502344.772753.56042.6019
114.7938-0.08380.19465.5798-2.57051.28010.6611-0.40080.323-0.64840.1147-0.3899-0.99540.0309-0.76140.9175-0.02940.2920.5156-0.04390.485541.284448.4126-2.4181
122.80140.61671.65851.26390.08924.1320.12350.24660.18720.06350.2966-0.2445-0.91591.108-0.4190.5113-0.17570.20590.6076-0.19750.459852.330541.4048-0.268
131.32850.43730.76792.13431.01696.1060.43180.05290.25670.01370.1881-0.1046-1.49290.1635-0.57540.8854-0.05220.26490.35890.0460.400442.410648.9837-7.125
143.74070.17970.981.66850.45182.18930.98290.7497-0.1541-0.0961-0.0722-0.2631-0.74460.1438-0.81860.34450.06630.17270.5323-0.02470.444342.96436.132-2.0684
157.6453-3.76140.46176.73931.60996.27910.72790.0732-0.4816-0.1321-0.08420.5045-0.5937-0.4196-0.6460.3480.04080.06040.34930.08980.294437.633336.492-3.6404
167.75293.52482.9846.3497-0.35914.03850.62170.8405-0.8515-0.32820.23-0.4112-0.4170.7579-0.82690.4195-0.06540.12060.4116-0.12610.313445.984936.4602-6.163
179.65096.2928-5.37782.0244-6.75265.2650.0546-0.765-0.5546-0.91070.0759-1.247-0.11041.6189-0.01670.646-0.1060.07231.0241-0.30930.627263.462233.95861.9482
185.8861-5.63030.64227.24560.46046.7550.18060.14790.80730.59310.1382-0.2816-0.9607-0.3891-0.30490.4130.02590.20170.47720.06080.533639.531842.21186.0769
196.3708-5.3671.22269.8148-1.45578.0090.3501-0.00550.13740.3373-0.2047-0.082-0.26-0.7493-0.2360.4188-0.12990.11410.5331-0.07590.289741.417340.00217.6334
203.5605-3.8788-1.80036.23182.92672.4937-0.1288-0.6124-0.16751.08530.23310.24640.37790.3491-0.06660.8372-0.11450.14750.55480.03940.388541.280127.28274.7218
212.1835-1.94991.01115.7833-1.90080.77190.2828-0.47010.13660.453-0.1441-0.1531-0.06230.1319-0.09490.8741-0.09030.18550.54820.03450.394136.176926.237571.4032
222.1261-0.5687-0.27651.95510.47881.75080.0607-0.2534-0.05790.3562-0.0977-0.07070.21250.36380.04070.5481-0.0480.10250.46050.01720.274445.889525.739662.985
234.1046-0.4978-1.23062.32711.31192.44310.1174-0.0755-0.11620.4852-0.08680.21860.0469-0.2021-0.00580.5505-0.05070.14570.2980.04420.278335.03630.126361.9251
242.01890.08720.62330.3383-0.01131.61070.00410.72240.11580.1661-0.07870.1655-0.0580.28630.09620.5112-0.08290.14690.3950.0030.356740.014730.951456.5506
252.03936.3259.73837.96224.80672.0564-0.28360.019-0.9681-0.32310.8342-0.895-1.02720.9961-0.51410.625-0.03110.12330.8316-0.13880.439257.105421.731456.4675
261.9815-5.04741.95827.92417.73391.992-0.9014-0.1052-0.50250.7868-0.00650.80272.4769-0.90220.92960.7449-0.13570.11440.4493-0.03030.454632.844420.237864.6976
279.6023-6.0692-7.3787.86396.36488.1768-0.5548-0.616-0.37670.84720.39650.67051.2791-0.02160.22530.6589-0.09570.10230.46870.06710.415134.562519.989566.2168
282.2785-1.0383-0.53972.6203-0.41911.402-0.0734-0.33070.32410.72280.0743-0.0543-0.59980.2277-0.00210.7375-0.14430.07910.4978-0.05990.343545.031854.880365.6495
296.0419-2.1498-1.40983.28070.32183.545-0.0143-0.15130.21250.34490.05520.2535-0.55270.1697-0.07010.5851-0.09540.10690.2809-0.05450.312938.35955.536556.5854
309.0493-3.90787.44836.5547-4.7219.47530.12361.1901-0.0227-0.0759-0.2821-0.3543-1.22781.92740.19150.9259-0.26380.20670.8573-0.1460.474848.693861.714259.0214
311.88871.7127-1.36122.855-3.17874.2220.5037-0.1397-0.37860.33880.22970.26581.4522-0.2672-0.51781.5265-0.2879-0.4860.54010.21550.799532.14536.1467-2.8417
320.4105-0.1091-0.63210.13550.58142.5219-0.0807-0.1802-1.0302-0.02540.22090.02991.73070.3146-0.33281.8904-0.032-0.59510.34770.10710.697637.484.6495-1.9241
337.5110.283-0.46192.7049-0.14040.0350.8225-0.0441-0.1536-0.4003-0.2311-0.14281.18610.2399-0.58281.17480.158-0.37780.4766-0.03380.601841.3978.7384-7.8851
340.3019-0.02160.34581.26660.09654.34780.3657-0.0991-0.2814-0.08780.33910.15491.8221-0.2325-0.60240.8817-0.1067-0.34830.4150.08050.540233.655114.7208-6.0533
353.5061-2.0263-1.42554.3305-0.00294.40470.67550.0148-0.1601-0.0880.0562-0.14330.94610.3635-0.63290.45750.0706-0.18190.3552-0.0660.362942.15722.1483-4.4798
368.71141.4899-0.91664.14810.58374.97010.59750.0025-0.1675-0.19360.28720.06430.768-0.3395-0.8630.5351-0.0235-0.20660.3350.04430.386536.786222.7841-7.8384
372.68030.2987-1.7139.29623.06316.0625-0.0859-0.5781-0.23880.30260.5883-0.33510.9879-0.238-0.45850.6394-0.2054-0.19420.37620.22640.361333.108817.14650.9575
382.28340.2921-1.3690.0361-0.22360.81580.49080.3084-0.96160.2417-0.3538-0.01751.8780.5373-0.10831.24670.2424-0.3690.5599-0.06570.651641.258611.25093.2119
393.55284.44712.92969.86393.59362.40870.1346-1.00280.40871.8798-0.05521.0166-0.3918-0.6821-0.19730.7221-0.01980.14520.5499-0.0310.361735.679636.835712.9177
401.88891.5474-1.01491.9752-1.20142.2425-0.33590.6078-0.5246-0.57010.4321-0.30212.073-0.6999-0.08851.4073-0.2547-0.43180.68960.06380.652731.69410.2291-36.2376
411.34780.3388-0.87541.5281-0.58812.86970.00160.3969-0.3922-0.63750.38680.40971.2658-0.7458-0.39820.9653-0.3269-0.33960.79830.1540.679626.950215.485-33.3642
422.9903-0.1375-1.22262.62820.51885.38890.10470.495-0.3203-0.43180.5427-0.01071.4074-0.2868-0.6310.7413-0.0751-0.27360.5209-0.00910.495536.510416.5213-31.123
430.57730.841-0.84983.45731.35174.4362-0.1643-0.54810.082-0.4271.0793-0.22370.39390.3069-0.83910.41670.0349-0.0930.59080.00260.396537.753126.4055-31.3479
444.5290.1724-1.8475.30822.6297.502-0.16810.07240.0071-0.11250.86820.7344-0.0947-0.2711-0.71340.3846-0.0444-0.0560.42450.15760.465433.834129.8251-29.431
453.27020.6314-1.92124.94760.11366.5222-0.40080.1444-0.253-0.12340.8063-0.66641.48931.158-0.38820.72980.062-0.17950.6686-0.20110.453646.310718.7276-32.1622
465.0594.6252-5.89966.5025-3.39618.383-0.57022.07580.5779-0.88191.46011.12590.278-1.1137-0.80190.8222-0.1216-0.28710.67210.0390.718530.340224.6509-38.195
479.58272.1731-7.3834.246-1.10426.7114-0.11870.43880.2359-1.30290.54370.61040.4086-1.0303-0.37331.0056-0.2392-0.40340.84570.13180.78832.852422.9771-40.7761
480.59330.2689-1.15520.9328-0.91012.99250.27480.32560.1237-0.38661.2943-0.5304-1.45881.6702-0.51971.3375-0.84740.76991.0933-0.22980.358454.5551.3774-29.3847
492.2259-0.40442.37821.4284-1.37583.811-0.0620.2534-0.132-0.3430.69950.1695-1.62710.2533-0.62321.0757-0.09760.29530.5113-0.05060.490339.943749.869-32.7325
501.3280.98870.82110.88780.44663.41650.23820.13170.0241-0.26720.5127-0.1191-1.03270.9787-0.70180.683-0.19150.28350.7054-0.20290.535449.317444.1052-25.1195
512.26161.73480.2291.5551-0.83935.4866-0.06650.0861-0.05160.15960.7156-0.99450.04521.0705-0.62950.4409-0.10880.12060.6419-0.31690.664349.031634.8818-28.8021
524.6394-3.7143.28964.7215-2.17394.41760.4830.0758-0.116-0.60260.28180.1113-0.9940.4385-0.74140.6034-0.13310.20130.496-0.05210.379142.946940.1305-25.5592
535.53712.31272.45953.8449-0.35465.17250.02340.7527-0.185-0.2030.422-0.1402-1.1030.5046-0.42091.0058-0.18990.23120.5034-0.02630.335144.163547.2498-33.7601
543.5907-3.11691.91924.4377-3.05083.10020.13810.57090.5472-1.2059-0.1558-0.1379-0.29631.23910.02810.9908-0.27620.35550.9794-0.23460.632549.916744.3603-38.256
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1CHAIN A AND (RESID 2:33)
2X-RAY DIFFRACTION2CHAIN A AND (RESID 34:152)
3X-RAY DIFFRACTION3CHAIN A AND (RESID 153:180)
4X-RAY DIFFRACTION4CHAIN A AND (RESID 181:205)
5X-RAY DIFFRACTION5CHAIN A AND (RESID 206:222)
6X-RAY DIFFRACTION6CHAIN B AND (RESID 3:152)
7X-RAY DIFFRACTION7CHAIN B AND (RESID 153:192)
8X-RAY DIFFRACTION8CHAIN B AND (RESID 193:230)
9X-RAY DIFFRACTION9CHAIN C AND (RESID 3:21)
10X-RAY DIFFRACTION10CHAIN C AND (RESID 22:47)
11X-RAY DIFFRACTION11CHAIN C AND (RESID 48:68)
12X-RAY DIFFRACTION12CHAIN C AND (RESID 69:111)
13X-RAY DIFFRACTION13CHAIN C AND (RESID 112:134)
14X-RAY DIFFRACTION14CHAIN C AND (RESID 135:152)
15X-RAY DIFFRACTION15CHAIN C AND (RESID 153:168)
16X-RAY DIFFRACTION16CHAIN C AND (RESID 169:180)
17X-RAY DIFFRACTION17CHAIN C AND (RESID 181:192)
18X-RAY DIFFRACTION18CHAIN C AND (RESID 193:205)
19X-RAY DIFFRACTION19CHAIN C AND (RESID 206:226)
20X-RAY DIFFRACTION20CHAIN D AND (RESID 3:33)
21X-RAY DIFFRACTION21CHAIN D AND (RESID 34:68)
22X-RAY DIFFRACTION22CHAIN D AND (RESID 69:111)
23X-RAY DIFFRACTION23CHAIN D AND (RESID 112:168)
24X-RAY DIFFRACTION24CHAIN D AND (RESID 169:180)
25X-RAY DIFFRACTION25CHAIN D AND (RESID 181:192)
26X-RAY DIFFRACTION26CHAIN D AND (RESID 193:205)
27X-RAY DIFFRACTION27CHAIN D AND (RESID 206:222)
28X-RAY DIFFRACTION28CHAIN E AND (RESID 3:134)
29X-RAY DIFFRACTION29CHAIN E AND (RESID 135:203)
30X-RAY DIFFRACTION30CHAIN E AND (RESID 204:222)
31X-RAY DIFFRACTION31CHAIN F AND (RESID 3:21)
32X-RAY DIFFRACTION32CHAIN F AND (RESID 22:44)
33X-RAY DIFFRACTION33CHAIN F AND (RESID 45:68)
34X-RAY DIFFRACTION34CHAIN F AND (RESID 69:134)
35X-RAY DIFFRACTION35CHAIN F AND (RESID 135:168)
36X-RAY DIFFRACTION36CHAIN F AND (RESID 169:180)
37X-RAY DIFFRACTION37CHAIN F AND (RESID 181:208)
38X-RAY DIFFRACTION38CHAIN F AND (RESID 209:219)
39X-RAY DIFFRACTION39CHAIN F AND (RESID 220:230)
40X-RAY DIFFRACTION40CHAIN G AND (RESID 3:21)
41X-RAY DIFFRACTION41CHAIN G AND (RESID 22:68)
42X-RAY DIFFRACTION42CHAIN G AND (RESID 69:134)
43X-RAY DIFFRACTION43CHAIN G AND (RESID 135:152)
44X-RAY DIFFRACTION44CHAIN G AND (RESID 153:168)
45X-RAY DIFFRACTION45CHAIN G AND (RESID 169:192)
46X-RAY DIFFRACTION46CHAIN G AND (RESID 193:208)
47X-RAY DIFFRACTION47CHAIN G AND (RESID 209:221)
48X-RAY DIFFRACTION48CHAIN H AND (RESID 3:68)
49X-RAY DIFFRACTION49CHAIN H AND (RESID 69:96)
50X-RAY DIFFRACTION50CHAIN H AND (RESID 97:152)
51X-RAY DIFFRACTION51CHAIN H AND (RESID 153:168)
52X-RAY DIFFRACTION52CHAIN H AND (RESID 169:180)
53X-RAY DIFFRACTION53CHAIN H AND (RESID 181:208)
54X-RAY DIFFRACTION54CHAIN H AND (RESID 209:221)

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