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- PDB-9ks5: The crystal structure of ABL1 in complex with K-CNBA-1 -

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Basic information

Entry
Database: PDB / ID: 9ks5
TitleThe crystal structure of ABL1 in complex with K-CNBA-1
ComponentsTyrosine-protein kinase ABL1
KeywordsUNKNOWN FUNCTION / ABL1 / K-CBA-1 / LYSINE MODIFICATION / IRREVERSIBLE COVALENT PROBE
Function / homology
Function and homology information


: / positive regulation of actin filament binding / negative regulation of ubiquitin-protein transferase activity / protein localization to cytoplasmic microtubule plus-end / DNA conformation change / DN4 thymocyte differentiation / response to epinephrine / activation of protein kinase C activity / phospholipase C-inhibiting G protein-coupled receptor signaling pathway / podocyte apoptotic process ...: / positive regulation of actin filament binding / negative regulation of ubiquitin-protein transferase activity / protein localization to cytoplasmic microtubule plus-end / DNA conformation change / DN4 thymocyte differentiation / response to epinephrine / activation of protein kinase C activity / phospholipase C-inhibiting G protein-coupled receptor signaling pathway / podocyte apoptotic process / delta-catenin binding / Role of ABL in ROBO-SLIT signaling / transitional one stage B cell differentiation / regulation of postsynaptic specialization assembly / regulation of modification of synaptic structure / nicotinate-nucleotide adenylyltransferase activity / cerebellum morphogenesis / neuroepithelial cell differentiation / B cell proliferation involved in immune response / positive regulation of extracellular matrix organization / positive regulation of Wnt signaling pathway, planar cell polarity pathway / microspike assembly / B-1 B cell homeostasis / neuropilin signaling pathway / neuropilin binding / bubble DNA binding / mitochondrial depolarization / regulation of cell motility / positive regulation of establishment of T cell polarity / activated T cell proliferation / cellular response to dopamine / positive regulation of blood vessel branching / proline-rich region binding / negative regulation of mitotic cell cycle / regulation of Cdc42 protein signal transduction / mitogen-activated protein kinase binding / regulation of hematopoietic stem cell differentiation / syntaxin binding / positive regulation of dendrite development / alpha-beta T cell differentiation / positive regulation of cell migration involved in sprouting angiogenesis / peptidyl-tyrosine autophosphorylation / regulation of axon extension / regulation of T cell differentiation / positive regulation of peptidyl-tyrosine phosphorylation / negative regulation of cell-cell adhesion / neuromuscular process controlling balance / HDR through Single Strand Annealing (SSA) / Myogenesis / positive regulation of osteoblast proliferation / platelet-derived growth factor receptor-beta signaling pathway / RUNX2 regulates osteoblast differentiation / positive regulation of vasoconstriction / Fc-gamma receptor signaling pathway involved in phagocytosis / vascular endothelial cell response to oscillatory fluid shear stress / regulation of endocytosis / Bergmann glial cell differentiation / regulation of microtubule polymerization / negative regulation of long-term synaptic potentiation / myoblast proliferation / associative learning / negative regulation of cellular senescence / actin monomer binding / signal transduction in response to DNA damage / positive regulation of focal adhesion assembly / canonical NF-kappaB signal transduction / negative regulation of BMP signaling pathway / RHO GTPases Activate WASPs and WAVEs / ephrin receptor signaling pathway / cardiac muscle cell proliferation / positive regulation of T cell migration / BMP signaling pathway / cellular response to transforming growth factor beta stimulus / endothelial cell migration / negative regulation of endothelial cell apoptotic process / negative regulation of double-strand break repair via homologous recombination / regulation of cell adhesion / mismatch repair / ephrin receptor binding / spleen development / four-way junction DNA binding / positive regulation of stress fiber assembly / ruffle / ERK1 and ERK2 cascade / phosphotyrosine residue binding / actin filament polymerization / positive regulation of substrate adhesion-dependent cell spreading / positive regulation of endothelial cell migration / positive regulation of interleukin-2 production / substrate adhesion-dependent cell spreading / positive regulation of mitotic cell cycle / SH2 domain binding / protein kinase C binding / peptidyl-tyrosine phosphorylation / response to endoplasmic reticulum stress / thymus development / Turbulent (oscillatory, disturbed) flow shear stress activates signaling by PIEZO1 and integrins in endothelial cells / positive regulation of release of sequestered calcium ion into cytosol / integrin-mediated signaling pathway / post-embryonic development
Similarity search - Function
F-actin binding / F-actin binding / F-actin binding domain (FABD) / Tyrosine-protein kinase ABL, SH2 domain / : / SH3 domain / SH2 domain / Src homology 2 (SH2) domain profile. / Src homology 2 domains / SH2 domain ...F-actin binding / F-actin binding / F-actin binding domain (FABD) / Tyrosine-protein kinase ABL, SH2 domain / : / SH3 domain / SH2 domain / Src homology 2 (SH2) domain profile. / Src homology 2 domains / SH2 domain / Src homology 3 domains / SH2 domain superfamily / SH3-like domain superfamily / Src homology 3 (SH3) domain profile. / SH3 domain / Tyrosine-protein kinase, catalytic domain / Tyrosine kinase, catalytic domain / Tyrosine protein kinases specific active-site signature. / Tyrosine-protein kinase, active site / Serine-threonine/tyrosine-protein kinase, catalytic domain / Protein tyrosine and serine/threonine kinase / Protein kinase, ATP binding site / Protein kinases ATP-binding region signature. / Protein kinase domain profile. / Protein kinase domain / Protein kinase-like domain superfamily
Similarity search - Domain/homology
Tyrosine-protein kinase ABL1
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.2 Å
AuthorsZhang, Z.M. / Duan, L.P.
Funding support1items
OrganizationGrant numberCountry
Not funded
CitationJournal: To Be Published
Title: O-Cyanobenzaldehydes (CNBA) Irreversibly Modify both Buried and Exposed Lysine Residues in Live Cells
Authors: Zhang, Z.M. / Duan, L.P.
History
DepositionNov 29, 2024Deposition site: PDBJ / Processing site: PDBC
Revision 1.0Dec 3, 2025Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Tyrosine-protein kinase ABL1
B: Tyrosine-protein kinase ABL1


Theoretical massNumber of molelcules
Total (without water)64,2672
Polymers64,2672
Non-polymers00
Water6,774376
1
A: Tyrosine-protein kinase ABL1


Theoretical massNumber of molelcules
Total (without water)32,1341
Polymers32,1341
Non-polymers00
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
B: Tyrosine-protein kinase ABL1


Theoretical massNumber of molelcules
Total (without water)32,1341
Polymers32,1341
Non-polymers00
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)106.48, 134.99, 58.0
Angle α, β, γ (deg.)90.0, 90.0, 90.0
Int Tables number18
Space group name H-MP21212
Space group name HallP22ab
Symmetry operation#1: x,y,z
#2: x+1/2,-y+1/2,-z
#3: -x+1/2,y+1/2,-z
#4: -x,-y,z

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Components

#1: Protein Tyrosine-protein kinase ABL1 / Abelson murine leukemia viral oncogene homolog 1 / Abelson tyrosine-protein kinase 1 / Proto- ...Abelson murine leukemia viral oncogene homolog 1 / Abelson tyrosine-protein kinase 1 / Proto-oncogene c-Abl / p150


Mass: 32133.697 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: ABL1, ABL, JTK7 / Production host: Escherichia coli (E. coli)
References: UniProt: P00519, non-specific protein-tyrosine kinase
#2: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 376 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestY
Has protein modificationY

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

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Sample preparation

CrystalDensity Matthews: 3.29 Å3/Da / Density % sol: 62.65 %
Crystal growTemperature: 298.15 K / Method: vapor diffusion, hanging drop / pH: 5
Details: 0.1 M Sodium citrate, 2% Tacinate (pH 5.0) and 16% w/v PEG 3350

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: CAMD / Beamline: GCPCC / Wavelength: 0.978 Å
DetectorType: DECTRIS PILATUS 300K / Detector: PIXEL / Date: Sep 15, 2024
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.978 Å / Relative weight: 1
ReflectionResolution: 2.2→32.2 Å / Num. obs: 43305 / % possible obs: 100 % / Redundancy: 9.8 % / Biso Wilson estimate: 8.78 Å2 / CC1/2: 0.998 / Rmerge(I) obs: 0.14 / Rpim(I) all: 0.048 / Rrim(I) all: 0.156 / Net I/σ(I): 15
Reflection shellResolution: 2.2→2.27 Å / Num. unique obs: 3714 / CC1/2: 0.836

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Processing

Software
NameVersionClassification
PHENIX1.9_1692+SVNrefinement
HKL-3000data reduction
HKL-3000data scaling
Cootmodel building
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.2→32.17 Å / SU ML: 0.226367327838 / Cross valid method: FREE R-VALUE / σ(F): 1.34351548402 / Phase error: 20.6713067915
RfactorNum. reflection% reflection
Rfree0.211556250836 1555 3.59579142097 %
Rwork0.170247344589 41690 -
obs0.171712121203 43245 99.9468429324 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso mean: 39.1266114246 Å2
Refinement stepCycle: LAST / Resolution: 2.2→32.17 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4321 0 76 376 4773
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.006346795291974517
X-RAY DIFFRACTIONf_angle_d0.8803048784626117
X-RAY DIFFRACTIONf_chiral_restr0.033813202477646
X-RAY DIFFRACTIONf_plane_restr0.00335285590363762
X-RAY DIFFRACTIONf_dihedral_angle_d13.63683908041657
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.2-2.2710.3219585196381390.2191374230523734X-RAY DIFFRACTION100
2.271-2.35220.262985249871390.2072667089673737X-RAY DIFFRACTION99.974206861
2.3522-2.44630.2274129331811400.1955227496323738X-RAY DIFFRACTION100
2.4463-2.55760.2605300595461390.1907110672453743X-RAY DIFFRACTION99.9742467165
2.5576-2.69240.2519603159271400.1960288656673749X-RAY DIFFRACTION99.9742930591
2.6924-2.8610.2328277352891400.194041256623752X-RAY DIFFRACTION99.9486389317
2.861-3.08170.2458562425731410.1882176158093784X-RAY DIFFRACTION99.9745287825
3.0817-3.39150.2258542752021410.1798976463853778X-RAY DIFFRACTION99.9489926039
3.3915-3.88160.2011345191420.1513825668223817X-RAY DIFFRACTION99.9747474747
3.8816-4.88760.1707343609991450.1318716804213851X-RAY DIFFRACTION99.9499749875
4.8876-32.170.1640080787891490.1595202545474007X-RAY DIFFRACTION99.7360211183
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)
13.010789689473.289095204313.947879283644.861305835956.04798688377.55789026035-0.201167300311-0.8437135351760.708639779290.604931240287-0.5255676895880.36836782688-0.153321854639-1.793835947060.7269160377150.4512023736060.108589740941-0.07336397556430.623531018406-0.1120484447570.364126312351-29.385500547221.13463142737.88134342064
22.87656431062-3.661273183730.2960038532969.290323080132.502849620155.5282462977-0.4329814971891.326159686620.0766380708735-0.030473250217-0.21756322329-0.518499132487-0.128494262934-0.1959936526490.6778658994760.241903538494-0.01784437119080.01137955508080.4047530701530.03152542500810.227941711466-12.708898945419.0806716635.53145020181
35.00711587587-2.941798495254.325383188734.53513993785-0.8160131894076.18922258624-0.249791045220.09936224094490.6609593664140.141946129634-0.31443128564-0.212062502613-0.4591832504870.1276565666190.5071584622210.2271194446120.01136438444710.02947853048250.249494825451-0.02353754229320.270063291013-22.353558567422.0682549788-0.64312130592
42.37932602383-0.9120859510710.2268006444231.46410556192-0.03501517543041.89196259769-0.152091437346-0.1311865362780.1089148884420.2103925566740.0761492528420.120811088134-0.114975828387-0.2377196960380.05628806876670.1708802232840.01442273782270.01714618727430.165733660365-0.004687983171610.166750384996-20.80242328319.84614542914-10.3710347734
54.51818477443-2.823512194020.5294286059934.55194927722-0.8603056543241.946576346550.07105691702980.1065802707460.388971118982-0.0917648722248-0.118225764629-0.160075315808-0.206037044572-0.1639600819930.05293719214320.1823839823970.02720425470590.0178037554760.1872033656430.009945299421880.12729903707-19.411026936717.1152612684-26.1304307877
63.171265355040.8120456827940.4759228187045.28851081196-5.996136211138.37680237014-0.1618360852110.12657458279-0.01974036882740.219010856352-0.100359531091-0.289741911625-0.3651711641830.4320289048720.3203474965910.1314076315540.04916245167110.01334598822910.2372676904640.003255300572420.188637563027-5.3751764762311.0404393656-25.9377727915
74.903334265984.842002759046.464207006894.89522145416.465418657968.58695588271-0.2380761858230.4794127092010.748701520021-0.691890796723-0.027554751020.104668607108-0.86238053267-0.1262090472840.1937737716330.3186319503330.04015173559880.01760282137550.2808925876550.06835434303130.236003108401-11.690388567719.8521108366-36.4813730977
83.4335897322-2.116870746561.825892374773.33258951335-4.566506924517.595416491070.05038238902590.117745279034-0.11815427482-0.367736319442-0.332136789244-0.5832362234060.5864772197850.4526769186770.2440136543830.2734544521340.06195430689070.001845666362120.257972437985-0.01773876188080.201843623335-9.774291175764.53206147593-32.984341083
99.52391943575-4.215907335465.286151026664.66637682228-5.710863838317.10222341003-0.11960544703-0.0556353117262-0.328809145725-0.4365173603480.4745466607290.02002175401870.4737276871230.125467278512-0.2560019998660.243452798821-0.003474976189150.03746850588850.273092997835-0.07667589859780.179355337073-17.00144289610.377763332476-30.9020912022
107.934150708781.66124613668-2.133078338664.9132951414-1.224944955073.355265319860.1740352466180.740090709372-0.0779231287241-0.576102182138-0.05646263447890.2932080141580.284380732943-0.601402960755-0.1358877347780.2139990021010.0327757024597-0.01299267130070.363379850246-0.04947454662870.149253116385-24.58366866859.78772609055-33.7398835868
116.35815697552-4.768777748563.029315650773.56485028912-2.403642534227.852578521170.1016773193640.108431228988-0.986958442444-0.03643742365070.09599809959920.5713976743161.20145025636-0.627404583061-0.2445872953590.311074253691-0.1092569693850.03771239059750.274384812168-0.03257583317750.337859755769-24.3640445247-4.69147918396-21.0505109014
124.053763892250.549278756888-2.827026011783.27305280083-0.6713471811357.9938426401-0.1999202004970.5484474023070.450129056049-0.3023438474680.329094243244-1.7382660181-1.079612435381.71225131025-0.09947510159720.572239886739-0.2817571630330.01843186388760.8253342159770.000418374415181.09296878847-9.7955052130357.1793689809-11.3179907863
133.347488792650.41526017715-0.6794534385033.851795553072.188988305857.074862646870.05621067080090.631875767657-0.12288242341-0.278551060951-0.238997384167-1.33309484433-0.4233192537530.7470225090850.09414652982860.423023891944-0.1254908979580.09456499367040.768460849109-0.10419588820.819262262882-14.69655487255.8234608918-16.4391431763
143.703567668450.0936877009405-3.746062329493.606155960093.736312961297.905522936840.6280422916410.3786254238770.217962851966-0.784207914626-0.175226121524-0.508129660366-1.27472908088-0.145169977501-0.4558813888820.4567443296540.02966410330770.1228798541530.4440549808380.03978510802690.546646700569-18.775005075362.7165020088-13.0258735345
154.134363706874.23897351055-6.089945845664.50535000036-6.329570033929.058367369760.5617985500690.3737970701211.256727968480.584688743828-0.1902017083010.919157368519-0.533693205739-0.695999349306-0.4277442326950.359215407354-0.0065348801035-0.03465212505160.405341313858-0.0636281893860.596159108334-23.737147155761.3407240555-3.32648494538
166.31368126324.9339083341-0.5952446227238.1857932308-0.5483998924526.19151976160.385608725605-0.2707389649040.1821896466140.334650517506-0.117372465671-0.86559780144-0.7828519591520.579577397016-0.2389287280070.3231650892090.0229494663932-0.07150831232910.339277193007-0.08075319686440.604818993083-18.876027944353.4637406871-5.1789317673
174.522941252812.600384493053.589177603647.972562031476.5873441186.3560579375-0.08591040710410.444132499332-0.210642456848-0.6776777771050.716193617679-1.13066347503-0.6109067424690.877712069053-0.7579743237240.28544870902-0.01592543383920.1367846527270.457834142051-0.02488323131070.509488946911-19.348709406751.1242846771-14.8995909437
189.65845856461-1.714013284521.563897881375.240184250772.551557637596.926796999360.1243273273630.8955981790240.0769056138691-1.41191047844-0.0230991556191-0.697893809571-0.1624732467890.385077434303-0.1531073455550.3008486159070.07146051771670.05582253699320.3940633156310.03976385511410.244188083593-32.559499410738.9274395172-25.815764688
194.084284040040.3208113748160.4670453531176.995222552685.469308041614.278632281-0.109813196046-0.165912758475-0.05322419055430.3031656084050.28085704597-0.3070010521780.2613925390930.372968867717-0.1634634767260.2398965658730.0687869125892-0.03512121732740.1856962276120.01323561825980.218122901059-34.229189532939.3381158284-8.1001200583
202.65483130371-2.97216503555-2.606955296488.596478900716.211203882724.594061462090.10292885636-0.1035832560230.280086002895-0.2132097451790.203542982094-0.467487011368-0.1001056903970.531945137195-0.3913835209730.2238920290730.0267143948841-0.007763436620940.259159014165-0.01282410478620.30115469819-31.814447317850.2037307409-7.20587324347
213.48917900313-4.70748481378-2.74973055046.613045651763.811269355032.20163673623-0.5504959760660.7825888715740.803369960781-1.03579826377-0.100326399711-0.549940842446-3.03079253860.5363162467130.632296801841.02426578768-0.128994463034-0.1587104971390.4044997381570.08031898041890.459846407965-40.555197731664.6735450336-8.83014343664
224.41618724417-4.83215091263-5.443196265327.607819087195.454292720236.80475064067-0.03253893960810.1942361704430.502222262027-0.5238260090380.0972593121144-0.0131753593833-0.4924529284260.167666996329-0.06827714743520.243675967968-0.0202735716581-0.03951087420190.1959531430270.02185938890410.250349878406-44.89282123254.94847413-15.5746075436
238.1755686999-0.3691475958226.081801259887.991867654351.880482010837.95636059167-0.567079883385-0.6720871830590.8810895203430.5184139808110.2482763083730.0707602166828-0.911252027992-0.2084136170460.2447844371960.4022138012970.0811440626351-0.02800574389350.276595081013-0.09076150108070.292559586836-47.279720771859.2582651307-5.36933667225
242.508843533760.428947740462-0.3896460275963.394408316252.405934478414.374386349830.08185453628630.3255332378910.142438831475-0.1640655992790.0973095657915-0.2560839072660.134467193570.0521733848734-0.1324994448440.1570672035080.07558423976090.003644201972450.204950118280.01743263764430.188980798671-43.974206010147.2883500234-17.0953575592
257.13962278358-3.66894987368-0.9623213480947.12229069950.655203328913.01624696890.09324987314050.4077998025270.0295770655982-0.430494044985-0.1997769331560.02895858023720.1404469085870.06932132700690.08123301957270.2021627812830.0267025009768-0.02818544048860.2099615342870.002295719577680.10153333562-53.760225845248.786312577-18.3384426057
268.32284564728-6.53398821714-5.706640322095.147317609614.487365641213.93673005583-0.07285494552040.137956705174-0.8310838631630.0277450956233-0.1287378815270.4118326288580.531138886093-0.4585741083820.07735661039810.289680735180.0257611832875-0.05289382847560.2511126728520.005069797922820.244847499161-49.57800009537.7724602444-12.6630086932
276.77413120639-5.76569215499-2.139230276076.776862691991.736827835511.91127248073-0.330003802011-0.218758758619-0.1666562983830.3944441719460.166483912130.02168013776560.1405974995570.0860084434120.1758449167990.2052582902570.0279933896967-0.01180659691170.1787608379550.02650626165640.146137941791-44.2606726438.5547004675-5.78516100365
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 233 through 241 )
2X-RAY DIFFRACTION2chain 'A' and (resid 242 through 250 )
3X-RAY DIFFRACTION3chain 'A' and (resid 252 through 292 )
4X-RAY DIFFRACTION4chain 'A' and (resid 293 through 392 )
5X-RAY DIFFRACTION5chain 'A' and (resid 394 through 433 )
6X-RAY DIFFRACTION6chain 'A' and (resid 434 through 444 )
7X-RAY DIFFRACTION7chain 'A' and (resid 445 through 453 )
8X-RAY DIFFRACTION8chain 'A' and (resid 454 through 465 )
9X-RAY DIFFRACTION9chain 'A' and (resid 466 through 475 )
10X-RAY DIFFRACTION10chain 'A' and (resid 476 through 485 )
11X-RAY DIFFRACTION11chain 'A' and (resid 486 through 500 )
12X-RAY DIFFRACTION12chain 'B' and (resid 233 through 247 )
13X-RAY DIFFRACTION13chain 'B' and (resid 248 through 265 )
14X-RAY DIFFRACTION14chain 'B' and (resid 266 through 279 )
15X-RAY DIFFRACTION15chain 'B' and (resid 280 through 291 )
16X-RAY DIFFRACTION16chain 'B' and (resid 292 through 310 )
17X-RAY DIFFRACTION17chain 'B' and (resid 311 through 322 )
18X-RAY DIFFRACTION18chain 'B' and (resid 323 through 336 )
19X-RAY DIFFRACTION19chain 'B' and (resid 337 through 356 )
20X-RAY DIFFRACTION20chain 'B' and (resid 357 through 392 )
21X-RAY DIFFRACTION21chain 'B' and (resid 394 through 402 )
22X-RAY DIFFRACTION22chain 'B' and (resid 403 through 407 )
23X-RAY DIFFRACTION23chain 'B' and (resid 408 through 417 )
24X-RAY DIFFRACTION24chain 'B' and (resid 418 through 444 )
25X-RAY DIFFRACTION25chain 'B' and (resid 445 through 465 )
26X-RAY DIFFRACTION26chain 'B' and (resid 466 through 475 )
27X-RAY DIFFRACTION27chain 'B' and (resid 476 through 500 )

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