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- PDB-8oz8: Crystal Structure of an Hydroxynitrile lyase variant (H96A) from ... -

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Basic information

Entry
Database: PDB / ID: 8oz8
TitleCrystal Structure of an Hydroxynitrile lyase variant (H96A) from Granulicella tundricola
ComponentsCupin 2 conserved barrel domain protein
KeywordsLYASE / Hydroxynitrile lyase / cupin / beta-barrel / ozonolysis / peroxidase
Function / homology
Function and homology information


(R)-mandelonitrile lyase / lyase activity / metal ion binding
Similarity search - Function
: / Cupin 2, conserved barrel / Cupin domain / RmlC-like cupin domain superfamily / RmlC-like jelly roll fold
Similarity search - Domain/homology
BROMIDE ION / : / 3-CARBOXY-N,N,N-TRIMETHYLPROPAN-1-AMINIUM / DI(HYDROXYETHYL)ETHER / 1-METHOXY-2-[2-(2-METHOXY-ETHOXY]-ETHANE / (R)-mandelonitrile lyase
Similarity search - Component
Biological speciesGranulicella tundricola (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.85 Å
AuthorsBento, I. / Coloma, J. / Hagedoorn, P.-L. / Hanefeld, U.
Funding support1items
OrganizationGrant numberCountry
Not funded
CitationJournal: Acs Catalysis / Year: 2023
Title: Can a Hydroxynitrile Lyase Catalyze an Oxidative Cleavage?
Authors: Coloma, J. / Hagedoorn, P.L. / Bento, I. / Hanefeld, U.
History
DepositionMay 8, 2023Deposition site: PDBE / Processing site: PDBE
Revision 1.0Apr 10, 2024Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Cupin 2 conserved barrel domain protein
B: Cupin 2 conserved barrel domain protein
C: Cupin 2 conserved barrel domain protein
D: Cupin 2 conserved barrel domain protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)58,54718
Polymers56,8034
Non-polymers1,74314
Water11,638646
1
A: Cupin 2 conserved barrel domain protein
B: Cupin 2 conserved barrel domain protein
C: Cupin 2 conserved barrel domain protein
D: Cupin 2 conserved barrel domain protein
hetero molecules

A: Cupin 2 conserved barrel domain protein
B: Cupin 2 conserved barrel domain protein
C: Cupin 2 conserved barrel domain protein
D: Cupin 2 conserved barrel domain protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)117,09336
Polymers113,6078
Non-polymers3,48628
Water1448
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation12_554x,x-y,-z-1/61
Buried area9940 Å2
ΔGint-80 kcal/mol
Surface area20120 Å2
2
A: Cupin 2 conserved barrel domain protein
B: Cupin 2 conserved barrel domain protein
C: Cupin 2 conserved barrel domain protein
D: Cupin 2 conserved barrel domain protein
hetero molecules

A: Cupin 2 conserved barrel domain protein
B: Cupin 2 conserved barrel domain protein
C: Cupin 2 conserved barrel domain protein
D: Cupin 2 conserved barrel domain protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)117,09336
Polymers113,6078
Non-polymers3,48628
Water1448
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation10_665-y+1,-x+1,-z+1/61
Buried area9660 Å2
ΔGint-81 kcal/mol
Surface area20430 Å2
Unit cell
Length a, b, c (Å)152.049, 152.049, 109.920
Angle α, β, γ (deg.)90.000, 90.000, 120.000
Int Tables number179
Space group name H-MP6522
Space group name HallP652(x,y,z+1/12)
Components on special symmetry positions
IDModelComponents
11A-433-

HOH

21A-444-

HOH

31B-471-

HOH

41C-438-

HOH

51C-452-

HOH

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Components

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Protein , 1 types, 4 molecules ABCD

#1: Protein
Cupin 2 conserved barrel domain protein


Mass: 14200.868 Da / Num. of mol.: 4 / Mutation: H96A
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Granulicella tundricola (bacteria) / Gene: AciX9_0562 / Production host: Escherichia coli (E. coli) / References: UniProt: E8WYN5

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Non-polymers , 7 types, 660 molecules

#2: Chemical ChemComp-2PE / NONAETHYLENE GLYCOL


Mass: 414.488 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C18H38O10 / Comment: precipitant*YM
#3: Chemical
ChemComp-NM2 / 3-CARBOXY-N,N,N-TRIMETHYLPROPAN-1-AMINIUM


Mass: 146.207 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C7H16NO2 / Feature type: SUBJECT OF INVESTIGATION
#4: Chemical
ChemComp-MN / MANGANESE (II) ION


Mass: 54.938 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Mn
#5: Chemical ChemComp-PG5 / 1-METHOXY-2-[2-(2-METHOXY-ETHOXY]-ETHANE


Mass: 178.226 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C8H18O4
#6: Chemical ChemComp-BR / BROMIDE ION


Mass: 79.904 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: Br
#7: Chemical ChemComp-PEG / DI(HYDROXYETHYL)ETHER


Mass: 106.120 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Formula: C4H10O3
#8: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 646 / Source method: isolated from a natural source / Formula: H2O

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Details

Has ligand of interestY

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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: 3.23 Å3/Da / Density % sol: 61.9 %
Crystal growTemperature: 292 K / Method: vapor diffusion, sitting drop / Details: KBr, PEG 2000 MME, natrium citrate

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: PETRA III, EMBL c/o DESY / Beamline: P13 (MX1) / Wavelength: 0.9763 Å
DetectorType: DECTRIS EIGER X 16M / Detector: PIXEL / Date: Dec 15, 2022
RadiationMonochromator: Oxford-FMB / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9763 Å / Relative weight: 1
ReflectionResolution: 1.85→131.68 Å / Num. obs: 64090 / % possible obs: 100 % / Redundancy: 11.1 % / Biso Wilson estimate: 25.22 Å2 / CC1/2: 0.995 / Rmerge(I) obs: 0.246 / Rpim(I) all: 0.111 / Rrim(I) all: 0.27 / Net I/σ(I): 8.7
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsNum. unique obsCC1/2Rpim(I) allRrim(I) all% possible all
9.06-131.689.60.0656560.9970.0280.07199.8
1.85-1.8911.52.25838970.5631.0012.472

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Processing

Software
NameVersionClassification
REFMAC1.20.1_4487refinement
PHENIX1.20.1_4487refinement
XDSdata reduction
Aimlessdata scaling
MOLREPphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.85→65.84 Å / SU ML: 0.1963 / Cross valid method: FREE R-VALUE / σ(F): 1.33 / Phase error: 18.6401
Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2
RfactorNum. reflection% reflection
Rfree0.1951 3232 5.05 %
Rwork0.1638 60806 -
obs0.1653 64038 99.96 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.1 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso mean: 30.78 Å2
Refinement stepCycle: LAST / Resolution: 1.85→65.84 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4008 0 72 646 4726
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00544225
X-RAY DIFFRACTIONf_angle_d0.75485763
X-RAY DIFFRACTIONf_chiral_restr0.0545596
X-RAY DIFFRACTIONf_plane_restr0.0075757
X-RAY DIFFRACTIONf_dihedral_angle_d10.4233594
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.85-1.880.33711370.29022624X-RAY DIFFRACTION99.96
1.88-1.910.32091310.27012592X-RAY DIFFRACTION99.93
1.91-1.940.26691240.24042616X-RAY DIFFRACTION100
1.94-1.970.24881520.22882598X-RAY DIFFRACTION99.96
1.97-2.010.26571650.20322569X-RAY DIFFRACTION99.96
2.01-2.050.23061700.1922568X-RAY DIFFRACTION99.96
2.05-2.090.19641400.1822614X-RAY DIFFRACTION100
2.09-2.130.20831410.17392609X-RAY DIFFRACTION99.96
2.13-2.180.20941470.17192609X-RAY DIFFRACTION99.96
2.18-2.240.22071390.17012609X-RAY DIFFRACTION100
2.24-2.30.21111260.1612623X-RAY DIFFRACTION99.96
2.3-2.370.23821490.16372617X-RAY DIFFRACTION100
2.37-2.440.22481260.15812643X-RAY DIFFRACTION99.89
2.44-2.530.20561410.15642622X-RAY DIFFRACTION100
2.53-2.630.20651360.16082629X-RAY DIFFRACTION99.96
2.63-2.750.20291210.15542665X-RAY DIFFRACTION100
2.75-2.90.19281380.15612648X-RAY DIFFRACTION100
2.9-3.080.19051320.16962674X-RAY DIFFRACTION100
3.08-3.310.1941340.15852660X-RAY DIFFRACTION99.96
3.31-3.650.18071370.15392682X-RAY DIFFRACTION100
3.65-4.180.16571660.142687X-RAY DIFFRACTION99.96
4.18-5.260.13061400.11862740X-RAY DIFFRACTION100
5.26-65.840.19261400.18432908X-RAY DIFFRACTION99.74
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)
14.685644831025.023608769336.556334057626.075288215536.41147265159.767040316720.303493643380.181456737466-0.3218467008280.2109847318770.20670598492-0.19489135321-0.0703636116680.330950403317-0.4836469450.2543655682680.00425058705841-0.04427605656670.2308002796920.01004232309770.33493117570442.97474383125.58669452278-3.74876829497
27.48602827064-4.65472576003-2.204516066743.411855382941.88605228331.18223582154-0.2908307491080.159871623637-0.6630769047790.3326925584540.00297989668710.2965183124530.415152313037-0.2549631243860.26904755020.259217256138-0.01682507695330.0279208475340.2467275796540.05911484417570.42316033521720.24232696289.750339194871.81403951491
34.94038576733-1.617220453493.929606618420.983945325029-0.7537733009414.733232848410.137939036697-0.135314540268-0.3954113079150.1051735554650.06827600795680.09988819111750.1588731841260.0779265428339-0.2376807438240.219685124714-0.002732307821120.002292900470230.2119003302590.05412413528590.27762886288834.286428044710.97070241343.96120389309
42.10936064097-0.8495746953720.4224853188550.875717253741-0.001732736715441.08828202346-0.0796239260882-0.111913785837-0.2113706350830.1220429669140.09184205792520.138052889320.0300277267336-0.0396149722988-0.05069849548170.2199945638080.007840631546960.0425811540770.2096389048770.04420574045540.2491785271924.782417476921.30766123415.10778805602
52.110755962660.3096734303030.1707267494760.9921999223540.01107232438640.167065374041-0.0337928505202-0.176520904011-0.05936422947250.1728289890490.08957194735650.214515961412-0.0420443218355-0.0346998881485-0.04550912318360.2440410539550.02652688567850.03792497677930.2094385136350.03615563176050.26759073552126.543233840524.09505132835.06080045277
66.15486608505-5.348105388864.60635460127.09518618917-3.226695872235.20743518314-0.0640143386825-0.943117595414-1.346149254250.1904047099190.319296137802-0.09233582756950.5781537161910.142807135586-0.1330768882830.3909665327630.0653482021540.1271944930180.3529634230790.1436006272870.59762222070228.231648214411.26781892915.3101893633
72.345307387063.01644195699-4.375581187483.97442369006-5.739488666328.28355769161-0.02289333737910.2096301113430.4705583001790.1755410024360.1648390865990.894919591823-0.243066550659-0.74840318326-0.4111919306590.2323596592770.03400912606030.01515664822280.314042692230.04736099356730.42578801559812.607858530723.30131402975.26484871645
81.180876502271.66399004108-1.19302257292.83889406504-2.649461119623.095422388480.0898596079325-0.1075008481980.02685162532640.4273318747590.01974293193630.406754033953-0.314476043706-0.0374178704824-0.1214341431930.325879267506-0.0022537693485-0.01866444534360.282398826133-0.02305463205130.23843742922341.32026629566.488367952123.7671313158
92.85785347521.896683327662.692472999445.061408546314.346169302824.267841807790.110010919223-0.3571508804660.04614658804130.253114919997-0.1528830665730.084171326372-0.0999708290581-0.3596914606390.05637534818490.281698173909-0.03623610462790.0251869915680.2771537073720.01842536204510.17184406926929.317916287747.871361105915.3270474925
101.08704752999-2.092436474790.1484805882438.41431324921.938372648861.40609365199-0.0374549766126-0.1852605384360.08220985164450.45515911306-0.007124538744540.2179346682590.00624014310691-0.1253380522650.02164524370430.230744906245-0.02607472233660.04651414805580.24124825356-0.0233055473450.21927621275232.868324323461.731995842915.3731794752
112.50842551306-0.1313490782140.3303673391131.74241254554-0.007008609275511.00277700356-0.01050909206240.0995803284830.129251702145-0.0518171476397-0.02983744132450.0715277578919-0.0344433013492-0.05768967849810.04397105757560.204430195719-0.006413816918910.004143349086830.202451064073-0.01502026186320.16738039728132.450932406154.56381402872.67008971677
121.527439362243.495087859842.079684475228.654466748034.432297466893.333585752080.124420555179-0.3922884065640.3969567435340.414456564306-0.5304501881830.883315246275-0.00403463697636-0.4567205378250.402965617150.259401903788-0.03064924069410.05922183251630.318127046535-0.05088760003440.29293920746621.263249169656.08228627669.53157912748
133.085689392930.3352477723382.750763176687.170676867633.049635362663.546604197930.127888855291.26770236296-1.19794340268-0.7982118379470.198812174773-0.2212879949580.2279584516240.336164984005-0.2915875592830.349593010381-0.005106028743650.006088529204340.385305822272-0.03909191037290.30169762344128.981410888840.8440846608-1.93858176513
145.329264963784.79667809427-5.249751601594.77011289067-4.417855265755.320189313950.0660567393201-0.325166420337-0.05024355137570.339767102392-0.007124598184910.0333128145262-0.05647775684490.13122079109-0.03489333803120.2545559148720.01539838292940.01574003907070.176730281942-0.01168883875980.22238752695734.7806357532.51573413164.50176863178
155.69373624374-4.63699793262-1.590118253685.644125549641.846154039513.85515600847-0.0308251533999-0.190724835910.4988827464140.3133155922930.044081528774-0.0689751698591-0.1411141757110.10874925713-0.003584625091090.201122025848-0.03185497476220.01417689287590.208506557451-0.02557195262880.22302099471557.635936350628.02512557650.776203842247
166.304662408250.696189205349-5.946044462012.35630142861-1.732608568416.060814865340.162095547416-0.431997437990.1403361337250.286844754654-0.004768598941410.133975487797-0.2978840769820.374471801787-0.221439928790.2146217848310.00488678806949-0.004602891739580.215840219368-0.03402093855160.1815057312144.703406589624.90823817186.14966980597
171.267498202510.30681660846-0.1084936899070.552032988789-0.5932980350622.00178448328-0.0260621384712-0.0442058864678-0.09096197396270.04006384225040.0785497542389-0.02283802910170.09270157518440.119231419478-0.05028041492170.1847602596610.0316467373915-0.002229355518690.186852988429-0.02089450316640.18424085459653.363536018615.9043835758-0.76644267006
180.8195890969170.327089180744-0.2808938436111.35296749518-0.3649745005952.349668192-0.00309405162179-0.085003068652-0.09123745079860.0769497304510.0194354765897-0.06100827137890.174742847030.133135934265-0.04729469644510.1643930413770.0193164343891-0.008005533466950.168441108997-0.01101702619950.16071492280251.492653818913.3272582206-1.80839311723
197.60056698553-1.38686053236-4.566678769090.9352061754311.194606015152.9200756092-0.169491548251-1.186514929590.2505186454160.6009311434880.2034674895340.0280553306968-0.318560141480.463629668341-0.02976301062540.3706502503780.04811039017270.03194447239580.374735107384-0.06066103220620.19821932564452.958537562719.903208740813.731240165
207.25492525310.6326544230710.7798166175533.692483063490.6198013989782.670067617130.05512445701440.316909709274-0.122588865767-0.259822095980.0103733993473-0.2861224963360.6631540576380.309088171664-0.1555216661640.2397982040930.04049130118860.008946137322660.239216354355-0.01222096064260.18019998758864.877320728914.3549123459-4.89738044645
215.651527182047.23643688725-7.24320448599.72517104354-8.686573614252.03099137566-0.179793493771-0.382100768456-0.4910449874010.103357431797-0.0686254869226-0.3734819087880.05197050471390.5409074451910.1979861854170.314207273076-0.0284206807594-0.006291156572310.30081547735-0.05999128531610.27292052699153.232906353247.5082278287-3.05085505074
220.8477788060440.505481489981-1.405063836884.92985071128-4.714569935136.864692885520.141488312387-0.0560951958565-0.511238574673-0.1425961483490.06460945217790.01150548391420.948039257092-0.0969772774311-0.2507582647780.352708609150.0114274674414-0.04017039805920.165605397833-0.02082277805550.27981616760548.878722833937.956099907218.0581855043
234.992143949372.50253177017-1.653325318821.46586203341-1.467334003184.97687992981-0.1688776042480.34837929362-0.533255533592-0.06780578693910.0564061482864-0.3626307480580.3405325765580.2449098692470.153238086660.2374113996580.0411275840556-0.006935070792790.1679520844-0.0341003740560.27197711224557.264325848644.72754503098.83961001242
242.782051478690.237935641074-0.09018185503092.05844454018-0.172429459922.037786598150.0599584360546-0.175765161823-0.2061748163340.2658026840130.00527421441555-0.2830417073140.1784671287180.160451398619-0.06024023504960.2664645836530.0145011572759-0.06818493128790.193992272490.008794832878140.20430840721456.436978607249.738785262322.6502431632
255.798645511995.32645388738-3.607063750476.39936156651-4.844925011343.8991957655-0.5890743744720.378984400526-1.0352592134-0.8469283405760.255868296887-1.115547254111.879646560420.8658726893610.3266861886120.6727877489160.163406512472-0.05669398855120.399970275522-0.1319515258160.55208010398864.973214031737.890169945515.9513936787
267.23342099911-2.566886149432.053374100848.44037444826-3.125077105772.2679674261-0.0319882802586-1.11559521381-0.5034462538691.239406578940.4176183690760.5191348195520.164017848844-0.473487437444-0.4024943997060.5385662709330.00473543529551-0.07677751022530.3390332324710.0986924281520.211684865950.927952801243.913428016233.0598762734
Refinement TLS group

Refine-ID: X-RAY DIFFRACTION

IDRefine TLS-IDSelection detailsAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11chain 'A' and (resid 1 through 10 )AA1 - 101 - 10
22chain 'A' and (resid 11 through 22 )AA11 - 2211 - 22
33chain 'A' and (resid 23 through 44 )AA23 - 4423 - 44
44chain 'A' and (resid 45 through 72 )AA45 - 7245 - 72
55chain 'A' and (resid 73 through 111 )AA73 - 11173 - 111
66chain 'A' and (resid 112 through 121 )AA112 - 121112 - 121
77chain 'A' and (resid 122 through 131 )AA122 - 131122 - 131
88chain 'B' and (resid 1 through 10 )BD1 - 101 - 10
99chain 'B' and (resid 11 through 22 )BD11 - 2211 - 22
1010chain 'B' and (resid 23 through 44 )BD23 - 4423 - 44
1111chain 'B' and (resid 45 through 111 )BD45 - 11145 - 111
1212chain 'B' and (resid 112 through 125 )BD112 - 125112 - 125
1313chain 'B' and (resid 126 through 131 )BD126 - 131126 - 131
1414chain 'C' and (resid 1 through 10 )CG1 - 101 - 10
1515chain 'C' and (resid 11 through 22 )CG11 - 2211 - 22
1616chain 'C' and (resid 23 through 44 )CG23 - 4423 - 44
1717chain 'C' and (resid 45 through 72 )CG45 - 7245 - 72
1818chain 'C' and (resid 73 through 111 )CG73 - 11173 - 111
1919chain 'C' and (resid 112 through 121 )CG112 - 121112 - 121
2020chain 'C' and (resid 122 through 131 )CG122 - 131122 - 131
2121chain 'D' and (resid 1 through 10 )DJ1 - 101 - 10
2222chain 'D' and (resid 11 through 22 )DJ11 - 2211 - 22
2323chain 'D' and (resid 23 through 44 )DJ23 - 4423 - 44
2424chain 'D' and (resid 45 through 111 )DJ45 - 11145 - 111
2525chain 'D' and (resid 112 through 121 )DJ112 - 121112 - 121
2626chain 'D' and (resid 122 through 131 )DJ122 - 131122 - 131

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