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- PDB-6wbt: 2.52 Angstrom Resolution Crystal Structure of 6-phospho-alpha-glu... -

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Basic information

Entry
Database: PDB / ID: 6wbt
Title2.52 Angstrom Resolution Crystal Structure of 6-phospho-alpha-glucosidase from Gut Microorganisms in Complex with NAD and Glucose-6-phosphate
Componentscellulase IBT99
KeywordsHYDROLASE / 6-phospho-alpha-glucosidase / Gut Microorganisms
Function / homology6-O-phosphono-alpha-D-glucopyranose / : / NICOTINAMIDE-ADENINE-DINUCLEOTIDE / PHOSPHATE ION
Function and homology information
Biological speciesmetagenomes (others)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.52 Å
AuthorsWu, R. / Kim, Y. / Endres, M. / Joachimiak, J.
CitationJournal: To Be Published
Title: 2.52 Angstrom Resolution Crystal Structure of 6-phospho-alpha-glucosidase from Gut Microorganisms in Complex with NAD and Glucose-6-phosphate
Authors: Wu, R. / Kim, Y. / Endres, M. / Joachimiak, J.
History
DepositionMar 27, 2020Deposition site: RCSB / Processing site: RCSB
Revision 1.0Mar 31, 2021Provider: repository / Type: Initial release
Revision 1.1Oct 18, 2023Group: Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession
Revision 1.2Nov 15, 2023Group: Data collection / Category: chem_comp_atom / chem_comp_bond / Item: _chem_comp_atom.atom_id / _chem_comp_bond.atom_id_2

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: cellulase IBT99
B: cellulase IBT99
C: cellulase IBT99
D: cellulase IBT99
hetero molecules


Theoretical massNumber of molelcules
Total (without water)209,44418
Polymers205,5384
Non-polymers3,90614
Water3,333185
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area17750 Å2
ΔGint-103 kcal/mol
Surface area61500 Å2
MethodPISA
Unit cell
Length a, b, c (Å)102.797, 219.724, 97.978
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number19
Space group name H-MP212121
Space group name HallP2ac2ab
Symmetry operation#1: x,y,z
#2: x+1/2,-y+1/2,-z
#3: -x,y+1/2,-z+1/2
#4: -x+1/2,-y,z+1/2

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Components

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Protein / Sugars , 2 types, 7 molecules ABCD

#1: Protein
cellulase IBT99


Mass: 51384.422 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) metagenomes (others) / Production host: Escherichia coli (E. coli) / Strain (production host): BL21DE3 / Variant (production host): gold
#3: Sugar ChemComp-G6P / 6-O-phosphono-alpha-D-glucopyranose / ALPHA-D-GLUCOSE-6-PHOSPHATE / 6-O-phosphono-alpha-D-glucose / 6-O-phosphono-D-glucose / 6-O-phosphono-glucose


Type: D-saccharide, alpha linking / Mass: 260.136 Da / Num. of mol.: 3
Source method: isolated from a genetically manipulated source
Formula: C6H13O9P / Feature type: SUBJECT OF INVESTIGATION
IdentifierTypeProgram
a-D-Glcp6PO3IUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0

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Non-polymers , 5 types, 196 molecules

#2: Chemical
ChemComp-MN / MANGANESE (II) ION


Mass: 54.938 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Formula: Mn
#4: Chemical
ChemComp-NAD / NICOTINAMIDE-ADENINE-DINUCLEOTIDE / Nicotinamide adenine dinucleotide


Mass: 663.425 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C21H27N7O14P2 / Feature type: SUBJECT OF INVESTIGATION / Comment: NAD*YM
#5: Chemical ChemComp-PO4 / PHOSPHATE ION / Phosphate


Mass: 94.971 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: PO4
#6: Chemical ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C2H6O2
#7: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 185 / 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: 2.74 Å3/Da / Density % sol: 55.13 %
Crystal growTemperature: 289 K / Method: vapor diffusion, sitting drop / pH: 6.2
Details: 0.2 M NaCl, 0.1 M Na/K phosphate pH 6.2, 20 %(w/v) PEG1000

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-ID / Wavelength: 0.9792 Å
DetectorType: DECTRIS PILATUS3 X 6M / Detector: PIXEL / Date: Oct 19, 2019
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9792 Å / Relative weight: 1
ReflectionResolution: 2.52→75.09 Å / Num. obs: 70393 / % possible obs: 92.7 % / Redundancy: 3.4 % / Biso Wilson estimate: 54.48 Å2 / CC1/2: 0.981 / Rmerge(I) obs: 0.121 / Net I/σ(I): 5
Reflection shellResolution: 2.52→2.56 Å / Redundancy: 3.4 % / Rmerge(I) obs: 0.978 / Mean I/σ(I) obs: 1 / Num. unique obs: 3335 / CC1/2: 0.402 / % possible all: 89.4

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Processing

Software
NameVersionClassification
PHENIX1.13_2998refinement
HKL-3000data reduction
HKL-3000data scaling
HKL-3000phasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 1U8X
Resolution: 2.52→70.92 Å / Cross valid method: FREE R-VALUE
RfactorNum. reflection% reflectionSelection details
Rfree0.266 3582 5 %random
Rwork0.227 ---
obs0.229 66728 92.87 %-
Displacement parametersBiso mean: 63.73 Å2
Refinement stepCycle: LAST / Resolution: 2.52→70.92 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms14066 0 181 185 14432
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.001714539
X-RAY DIFFRACTIONf_angle_d0.489319691
X-RAY DIFFRACTIONf_chiral_restr0.04172196
X-RAY DIFFRACTIONf_plane_restr0.00332487
X-RAY DIFFRACTIONf_dihedral_angle_d20.85245593
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.52-2.550.34231060.33642619X-RAY DIFFRACTION95.38
2.55-2.590.36351330.35332617X-RAY DIFFRACTION94.86
2.59-2.630.38911270.33882596X-RAY DIFFRACTION95.54
2.63-2.660.44551260.3262621X-RAY DIFFRACTION95.08
2.66-2.710.33841500.32822572X-RAY DIFFRACTION94.81
2.71-2.750.33781330.29772579X-RAY DIFFRACTION94.86
2.75-2.80.35021320.30532598X-RAY DIFFRACTION94.2
2.8-2.850.33291380.29072602X-RAY DIFFRACTION95.14
2.85-2.90.32861240.28892504X-RAY DIFFRACTION91.41
2.9-2.960.29921240.29262589X-RAY DIFFRACTION93.49
2.96-3.030.31531270.27712596X-RAY DIFFRACTION94.58
3.03-3.10.34631450.28192568X-RAY DIFFRACTION94.04
3.1-3.170.29181520.29022541X-RAY DIFFRACTION93.25
3.17-3.260.34971610.292543X-RAY DIFFRACTION93.34
3.26-3.360.31191320.26872552X-RAY DIFFRACTION93.1
3.36-3.470.32151490.24672590X-RAY DIFFRACTION93.58
3.47-3.590.27361450.23152465X-RAY DIFFRACTION90.47
3.59-3.730.27271490.21582570X-RAY DIFFRACTION93.37
3.73-3.90.24561460.20532563X-RAY DIFFRACTION92.87
3.9-4.110.25151320.18832539X-RAY DIFFRACTION91.57
4.11-4.370.24241550.17422506X-RAY DIFFRACTION91.44
4.37-4.70.21941230.17382551X-RAY DIFFRACTION90.43
4.7-5.180.21161510.17082580X-RAY DIFFRACTION92.7
5.18-5.920.2261560.20352535X-RAY DIFFRACTION90.61
5.92-7.460.26351350.21122506X-RAY DIFFRACTION87.86
7.46-70.920.16561310.16562626X-RAY DIFFRACTION87.58
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)
10.806907179814-0.428520585321-0.3768809146680.354233426016-0.1127750110120.339671645316-0.02728915717870.5100784095410.0273425626214-0.084853370326-0.07171857772950.3809941130040.350545594132-0.527829488570.1564919014360.541721863606-0.185257314836-0.007151818941190.946600659449-0.0552020670110.857942679028-29.12693778784.0602219496221.462350467
21.44266697266-0.321090190702-0.8487172552230.9799373370260.7658055188692.42756902611-0.004917179402440.0664272203345-0.269965826164-0.0539794276113-0.02853550343840.11114730479-0.0425344727212-0.4452162214140.06649224008270.2575961980830.0295216696321-0.05030110220880.4489558829570.02229737010110.370816104779-14.075632699719.12879353217.8140079586
30.941584110212-0.512811811912-0.4839894914811.587469615030.7155629507111.60247886001-0.0533293327055-0.0521179793662-0.104670410479-0.02994988034460.0764100351157-0.09215390093250.1306026420190.290780502543-0.04586368214070.3249952312650.0440756671615-0.01458383249560.458190569885-0.01920581852820.47036376847327.75737110181.217037396869.88614487256
43.891957867320.676437052756-0.2776058224412.287165861530.2127813511294.5040285771-0.217765313960.533456453636-0.437789087905-0.5700317572770.279213880443-0.10351686470.3438897303830.252890603882-0.07459435026920.5385610617930.0086535924258-0.007976190307690.485072309813-0.1038903150280.48405921021910.698397528111.5544032588-8.48007003899
50.653684475808-0.990875523956-0.6084574896350.6404485307630.1207334559340.7176976690140.02218920580380.150302100031-0.222489940676-0.0562748978188-0.03571659351920.009586791221840.146352877545-0.01692659722970.0136395047880.3713522254250.00467586527973-0.02346757754370.403484571871-0.03093002198210.50349601690412.99215764144.452903959978.17273155568
61.063521376260.3680470484220.1886760724161.653026856760.3757994678440.6014291425210.01865947310380.1874827878410.0931867533802-0.230126464930.0609355294367-0.170632819103-0.2948856823540.24797215777-0.07464503536510.605896100544-0.1498417119870.03019922971320.4725104217690.04011947108840.34543465915822.19992040455.8012898292-5.61063229945
70.740771894375-0.334205037947-0.3041794637331.41142754357-0.06314910974892.59741267040.0368081632950.124054847744-0.117775236541-0.129447134170.0696504982111-0.0594828754689-0.368351714356-0.145366174893-0.1022456422950.553777832116-0.1117962292950.02233262912490.3931109846520.01308045892830.39591952707412.468748070649.34335322795.12212461861
82.86361156879-1.534041002570.3236887980821.84202152608-0.3558129779242.938491592810.07057801678280.149965100622-0.166951379888-0.175049481759-0.0856785886229-0.499101451418-0.2956032662010.7478297470250.008022792041790.512156387111-0.1732835310930.01900407232660.561449984846-0.03946517219390.5206910628635.637334843141.419949846115.9586426546
90.157080188941-0.7810045430540.4495331384133.98062652967-2.074867226781.07759404956-0.159011013433-0.005073580942670.1153327883990.4138625049110.139319872152-0.198993685565-0.6253323586920.1578254826350.06260603195020.753941088069-0.229776166247-0.008472049059190.712241332642-0.04991809400260.56976040698628.421416357854.183459808222.5192608798
100.8521568804450.220352657394-0.6795158707261.19018291286-1.109014351280.5503273934960.01735071753450.03020763180030.03436541215150.0390997966185-0.0134983478196-0.0178560747769-0.3346047862050.170198833893-0.01774003828440.507434932014-0.105236526518-0.0145120355440.343037810222-0.01313729836970.30655401801417.048145805349.117286756611.1555124121
111.996958049740.672092902272-0.7650003197730.9889774251920.1094379162172.572620711690.100170082428-0.516398340372-0.167838375760.193600127647-0.128850178423-0.061032454958-0.2215626120320.2134042811620.02146792970070.4157323941280.00611994340791-0.04485462471850.5639640492830.08645736687620.3035583612354.7835552933633.460222982957.9907192751
123.892902945360.677342619204-0.9924135820792.64569482782-0.2334801196983.404177240520.355345762045-0.6607491068280.3553134160890.410636382610.007680607194510.382858857184-0.6968379978110.0874426051886-0.3444529778750.8155134068660.05402975460530.103781655180.5012501168640.01727314936530.447043446485-6.5147272238551.579884403241.8819711817
131.155566796980.566734442364-0.0007812962602380.05339801866280.2694854291772.53261828640.135999671342-0.2752599087440.04605770754620.190937802256-0.0525555198367-0.0241744501393-0.4945452354070.0775732755705-0.08362561964830.498437659847-0.02077782856130.005931640491590.3590230592030.02196820007860.3720483597266.6793202148440.334212995946.9972462462
143.75916069014-0.600962199875-0.8035956699764.809961873910.09423142276673.528436563940.106450077076-0.183424433758-0.6102027077620.108620418746-0.2722938074110.622215857940.135798108144-1.281424521370.173501935710.3583853621620.101967811563-0.01403524654751.00823185910.06410827512490.64949803633-37.24871938223.778637725528.1197740117
151.586068810730.118688641942-0.2405506956311.1457319475-0.0114884962081.986865435940.0808445682920.321108083612-0.2164483039570.0175448911103-0.07237069868190.332646100843-0.133214450037-1.004135281490.01349065539120.3649029970840.0722420178479-0.04041515295210.9434964455380.01302250476010.501632549049-30.926776484222.635676663818.6559590658
162.827084385450.398545249869-0.5692121036654.28762858154-0.3170091947953.9036509748-0.133574294987-0.0999449161349-0.4051592803580.381332222570.121514652490.09288776508220.531861316083-0.2969475971850.04438043990580.395457858087-0.02364331845910.03686162835610.5401220484230.09593036601070.561208252745-16.894614850110.98829580335.8906950828
173.62601670834-1.04425125616-2.584885400185.688952117080.84912941646.3535842296-0.297994877985-0.231726534166-0.889092427590.118662317738-0.2729721314650.3144205469070.5876316039520.03911497538030.541999179970.56532486017-0.07759598968340.06520467168920.4717739943150.07204715268920.649859400254-14.75568520920.66571851613536.5890437037
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'D' and (resid 256 through 319 )
2X-RAY DIFFRACTION2chain 'D' and (resid 320 through 443 )
3X-RAY DIFFRACTION3chain 'A' and (resid 2 through 174 )
4X-RAY DIFFRACTION4chain 'A' and (resid 175 through 256 )
5X-RAY DIFFRACTION5chain 'A' and (resid 257 through 443 )
6X-RAY DIFFRACTION6chain 'B' and (resid 2 through 104 )
7X-RAY DIFFRACTION7chain 'B' and (resid 105 through 174 )
8X-RAY DIFFRACTION8chain 'B' and (resid 175 through 243 )
9X-RAY DIFFRACTION9chain 'B' and (resid 244 through 293 )
10X-RAY DIFFRACTION10chain 'B' and (resid 294 through 443 )
11X-RAY DIFFRACTION11chain 'C' and (resid 3 through 174 )
12X-RAY DIFFRACTION12chain 'C' and (resid 175 through 278 )
13X-RAY DIFFRACTION13chain 'C' and (resid 279 through 443 )
14X-RAY DIFFRACTION14chain 'D' and (resid 2 through 26 )
15X-RAY DIFFRACTION15chain 'D' and (resid 27 through 174 )
16X-RAY DIFFRACTION16chain 'D' and (resid 175 through 210 )
17X-RAY DIFFRACTION17chain 'D' and (resid 211 through 255 )

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