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- PDB-3dwf: Crystal Structure of the Guinea Pig 11beta-Hydroxysteroid Dehydro... -

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Basic information

Entry
Database: PDB / ID: 3dwf
TitleCrystal Structure of the Guinea Pig 11beta-Hydroxysteroid Dehydrogenase Type 1 Mutant F278E
Components11-beta-hydroxysteroid dehydrogenase 1
KeywordsOXIDOREDUCTASE / Rossmann Fold / short chain dehydrogenase reductase / SDR / Endoplasmic reticulum / Glycoprotein / Lipid metabolism / Membrane / NADP / Signal-anchor / Steroid metabolism / Transmembrane
Function / homology
Function and homology information


11beta-hydroxysteroid dehydrogenase / 11-beta-hydroxysteroid dehydrogenase (NADP+) activity / cortisol dehydrogenase activity / 7beta-hydroxysteroid dehydrogenase (NADP+) / 7-beta-hydroxysteroid dehydrogenase (NADP+) activity / steroid metabolic process / lung development / NADP binding / endoplasmic reticulum membrane / protein homodimerization activity
Similarity search - Function
short chain dehydrogenase / Short-chain dehydrogenase/reductase, conserved site / Short-chain dehydrogenases/reductases family signature. / Short-chain dehydrogenase/reductase SDR / NAD(P)-binding Rossmann-like Domain / NAD(P)-binding domain superfamily / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE / 11-beta-hydroxysteroid dehydrogenase 1
Similarity search - Component
Biological speciesCavia porcellus (domestic guinea pig)
MethodX-RAY DIFFRACTION / SYNCHROTRON / Resolution: 2.2 Å
AuthorsLawson, A.J. / Ride, J.P. / White, S.A. / Walker, E.A.
CitationJournal: Protein Sci. / Year: 2009
Title: Mutations of key hydrophobic surface residues of 11 beta-hydroxysteroid dehydrogenase type 1 increase solubility and monodispersity in a bacterial expression system
Authors: Lawson, A.J. / Walker, E.A. / White, S.A. / Dafforn, T.R. / Stewart, P.M. / Ride, J.P.
History
DepositionJul 22, 2008Deposition site: RCSB / Processing site: RCSB
Revision 1.0Aug 11, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Non-polymer description / Version format compliance
Revision 1.2Oct 20, 2021Group: Database references / Derived calculations / Category: database_2 / struct_ref_seq_dif / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ref_seq_dif.details / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id
Revision 1.3Feb 21, 2024Group: Data collection / Category: chem_comp_atom / chem_comp_bond

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: 11-beta-hydroxysteroid dehydrogenase 1
B: 11-beta-hydroxysteroid dehydrogenase 1
C: 11-beta-hydroxysteroid dehydrogenase 1
D: 11-beta-hydroxysteroid dehydrogenase 1
hetero molecules


Theoretical massNumber of molelcules
Total (without water)125,63521
Polymers121,4454
Non-polymers4,19117
Water8,143452
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
A: 11-beta-hydroxysteroid dehydrogenase 1
B: 11-beta-hydroxysteroid dehydrogenase 1
hetero molecules


Theoretical massNumber of molelcules
Total (without water)62,6789
Polymers60,7222
Non-polymers1,9557
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area10160 Å2
ΔGint-97 kcal/mol
Surface area21300 Å2
MethodPISA
3
C: 11-beta-hydroxysteroid dehydrogenase 1
D: 11-beta-hydroxysteroid dehydrogenase 1
hetero molecules


Theoretical massNumber of molelcules
Total (without water)62,95812
Polymers60,7222
Non-polymers2,23510
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area10080 Å2
ΔGint-102 kcal/mol
Surface area21850 Å2
MethodPISA
Unit cell
Length a, b, c (Å)78.080, 85.920, 176.280
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number19
Space group name H-MP212121

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Components

#1: Protein
11-beta-hydroxysteroid dehydrogenase 1 / Corticosteroid 11-beta-dehydrogenase isozyme 1 / 11-DH / 11-beta-HSD1


Mass: 30361.188 Da / Num. of mol.: 4 / Mutation: F278E
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Cavia porcellus (domestic guinea pig) / Gene: HSD11B1 / Plasmid: pET28b(+) / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: Q6QLL4, 11beta-hydroxysteroid dehydrogenase
#2: Chemical
ChemComp-NAP / NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE / 2'-MONOPHOSPHOADENOSINE 5'-DIPHOSPHORIBOSE / Nicotinamide adenine dinucleotide phosphate


Mass: 743.405 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C21H28N7O17P3
#3: Chemical
ChemComp-SO4 / SULFATE ION / Sulfate


Mass: 96.063 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: SO4
#4: Chemical
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL / Glycerol


Mass: 92.094 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C3H8O3
#5: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 452 / Source method: isolated from a natural source / Formula: H2O

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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: 2.43 Å3/Da / Density % sol: 49.47 %
Crystal growTemperature: 291 K / Method: vapor diffusion / pH: 8
Details: 0.1M Tris-HCl, 1.75M ammonium sulphate and 5% glycerol, pH 8, Vapor diffusion, temperature 291K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ESRF / Beamline: ID14-1 / Wavelength: 0.933 Å
DetectorType: ADSC QUANTUM 4 / Detector: CCD / Date: Jul 15, 2007
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.933 Å / Relative weight: 1
ReflectionResolution: 2.2→34.68 Å / Num. obs: 60967 / Biso Wilson estimate: 34.468 Å2

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Processing

Software
NameVersionClassificationNB
PHENIXrefinement
PDB_EXTRACT3.005data extraction
RefinementResolution: 2.2→34.68 Å / FOM work R set: 0.834 / Stereochemistry target values: ml
RfactorNum. reflection% reflection
Rfree0.255 3085 5.07 %
Rwork0.2 --
obs-60836 99.77 %
Solvent computationBsol: 63.469 Å2 / ksol: 0.375 e/Å3
Displacement parametersBiso max: 180.64 Å2 / Biso mean: 43.75 Å2 / Biso min: 10.48 Å2
Baniso -1Baniso -2Baniso -3
1--0.133 Å20 Å20 Å2
2--0.029 Å20 Å2
3---0.104 Å2
Refinement stepCycle: LAST / Resolution: 2.2→34.68 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms8436 0 265 452 9153
Refine LS restraints
Refine-IDTypeDev idealWeight
X-RAY DIFFRACTIONf_angle_d0.9231
X-RAY DIFFRACTIONf_bond_d0.0051
X-RAY DIFFRACTIONf_chiral_restr0.0671
X-RAY DIFFRACTIONf_dihedral_angle_d16.811
X-RAY DIFFRACTIONf_plane_restr0.0031
X-RAY DIFFRACTIONf_nbd_refined4.121
LS refinement shell
Resolution (Å)Rfactor RworkNum. reflection RworkRefine-IDTotal num. of bins used% reflection obs (%)
2.2-2.2060.334458X-RAY DIFFRACTION11688
2.206-2.2130.309444X-RAY DIFFRACTION11691
2.213-2.2190.305494X-RAY DIFFRACTION11689
2.219-2.2260.309463X-RAY DIFFRACTION11693
2.226-2.2330.3485X-RAY DIFFRACTION11692
2.233-2.2390.298483X-RAY DIFFRACTION11694
2.239-2.2460.295485X-RAY DIFFRACTION11693
2.246-2.2530.283474X-RAY DIFFRACTION11695
2.253-2.260.282511X-RAY DIFFRACTION11696
2.26-2.2670.279483X-RAY DIFFRACTION11694
2.267-2.2740.28498X-RAY DIFFRACTION11696
2.274-2.2820.259495X-RAY DIFFRACTION11696
2.282-2.2890.266482X-RAY DIFFRACTION11695
2.289-2.2960.264508X-RAY DIFFRACTION11694
2.296-2.3040.271463X-RAY DIFFRACTION11695
2.304-2.3120.269518X-RAY DIFFRACTION11695
2.312-2.3190.249504X-RAY DIFFRACTION11695
2.319-2.3270.265490X-RAY DIFFRACTION11696
2.327-2.3350.249467X-RAY DIFFRACTION11695
2.335-2.3430.254513X-RAY DIFFRACTION11697
2.343-2.3510.245501X-RAY DIFFRACTION11697
2.351-2.360.256493X-RAY DIFFRACTION11695
2.36-2.3680.233526X-RAY DIFFRACTION11696
2.368-2.3770.243480X-RAY DIFFRACTION11696
2.377-2.3850.259486X-RAY DIFFRACTION11695
2.385-2.3940.257473X-RAY DIFFRACTION11693
2.394-2.4030.25487X-RAY DIFFRACTION11694
2.403-2.4120.239500X-RAY DIFFRACTION11695
2.412-2.4210.23495X-RAY DIFFRACTION11694
2.421-2.4310.261483X-RAY DIFFRACTION11695
2.431-2.440.252518X-RAY DIFFRACTION11694
2.44-2.450.238483X-RAY DIFFRACTION11696
2.45-2.460.239488X-RAY DIFFRACTION11694
2.46-2.470.217497X-RAY DIFFRACTION11697
2.47-2.480.238498X-RAY DIFFRACTION11695
2.48-2.490.226472X-RAY DIFFRACTION11694
2.49-2.50.225479X-RAY DIFFRACTION11694
2.5-2.5110.221537X-RAY DIFFRACTION11695
2.511-2.5220.226490X-RAY DIFFRACTION11694
2.522-2.5330.222489X-RAY DIFFRACTION11694
2.533-2.5440.197472X-RAY DIFFRACTION11694
2.544-2.5560.207484X-RAY DIFFRACTION11696
2.556-2.5670.203498X-RAY DIFFRACTION11694
2.567-2.5790.214508X-RAY DIFFRACTION11694
2.579-2.5910.19492X-RAY DIFFRACTION11696
2.591-2.6030.192511X-RAY DIFFRACTION11694
2.603-2.6160.19496X-RAY DIFFRACTION11696
2.616-2.6290.195467X-RAY DIFFRACTION11695
2.629-2.6420.186507X-RAY DIFFRACTION11695
2.642-2.6550.19485X-RAY DIFFRACTION11696
2.655-2.6680.198500X-RAY DIFFRACTION11695
2.668-2.6820.201515X-RAY DIFFRACTION11695
2.682-2.6960.179490X-RAY DIFFRACTION11693
2.696-2.7110.196488X-RAY DIFFRACTION11695
2.711-2.7260.186484X-RAY DIFFRACTION11696
2.726-2.740.182491X-RAY DIFFRACTION11694
2.74-2.7560.178503X-RAY DIFFRACTION11695
2.756-2.7720.182505X-RAY DIFFRACTION11694
2.772-2.7880.202470X-RAY DIFFRACTION11694
2.788-2.8040.19497X-RAY DIFFRACTION11693
2.804-2.8210.193489X-RAY DIFFRACTION11696
2.821-2.8380.182515X-RAY DIFFRACTION11696
2.838-2.8560.179484X-RAY DIFFRACTION11695
2.856-2.8740.197510X-RAY DIFFRACTION11696
2.874-2.8930.193499X-RAY DIFFRACTION11694
2.893-2.9120.198481X-RAY DIFFRACTION11696
2.912-2.9320.187489X-RAY DIFFRACTION11693
2.932-2.9520.19503X-RAY DIFFRACTION11697
2.952-2.9730.205516X-RAY DIFFRACTION11694
2.973-2.9940.214478X-RAY DIFFRACTION11694
2.994-3.0160.211500X-RAY DIFFRACTION11697
3.016-3.0390.193502X-RAY DIFFRACTION11695
3.039-3.0620.196510X-RAY DIFFRACTION11695
3.062-3.0860.201479X-RAY DIFFRACTION11696
3.086-3.1110.204507X-RAY DIFFRACTION11696
3.111-3.1370.205499X-RAY DIFFRACTION11695
3.137-3.1630.212487X-RAY DIFFRACTION11695
3.163-3.1910.201509X-RAY DIFFRACTION11695
3.191-3.2190.192499X-RAY DIFFRACTION11694
3.219-3.2490.209496X-RAY DIFFRACTION11694
3.249-3.280.196498X-RAY DIFFRACTION11695
3.28-3.3110.179510X-RAY DIFFRACTION11695
3.311-3.3440.193474X-RAY DIFFRACTION11696
3.344-3.3790.194506X-RAY DIFFRACTION11695
3.379-3.4150.206535X-RAY DIFFRACTION11695
3.415-3.4520.201475X-RAY DIFFRACTION11696
3.452-3.4910.195506X-RAY DIFFRACTION11696
3.491-3.5330.187494X-RAY DIFFRACTION11695
3.533-3.5760.181512X-RAY DIFFRACTION11694
3.576-3.6210.171486X-RAY DIFFRACTION11695
3.621-3.6680.171474X-RAY DIFFRACTION11693
3.668-3.7180.181539X-RAY DIFFRACTION11696
3.718-3.7720.174491X-RAY DIFFRACTION11694
3.772-3.8280.168496X-RAY DIFFRACTION11696
3.828-3.8880.164511X-RAY DIFFRACTION11696
3.888-3.9510.172505X-RAY DIFFRACTION11695
3.951-4.0190.181499X-RAY DIFFRACTION11693
4.019-4.0920.169520X-RAY DIFFRACTION11695
4.092-4.1710.167490X-RAY DIFFRACTION11695
4.171-4.2560.169497X-RAY DIFFRACTION11695
4.256-4.3480.162525X-RAY DIFFRACTION11697
4.348-4.4490.161494X-RAY DIFFRACTION11694
4.449-4.560.161515X-RAY DIFFRACTION11695
4.56-4.6830.153514X-RAY DIFFRACTION11696
4.683-4.8210.159518X-RAY DIFFRACTION11694
4.821-4.9760.164492X-RAY DIFFRACTION11695
4.976-5.1530.169518X-RAY DIFFRACTION11695
5.153-5.3590.188512X-RAY DIFFRACTION11695
5.359-5.6020.203516X-RAY DIFFRACTION11695
5.602-5.8960.199510X-RAY DIFFRACTION11695
5.896-6.2630.199522X-RAY DIFFRACTION11696
6.263-6.7430.201519X-RAY DIFFRACTION11694
6.743-7.4160.191531X-RAY DIFFRACTION11696
7.416-8.4750.185530X-RAY DIFFRACTION11694
8.475-10.6270.178535X-RAY DIFFRACTION11694
10.627-34.6840.242574X-RAY DIFFRACTION11695
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)
1-0.0556-0.0957-0.14112.04460.18730.8369-0.0809-0.07780.03220.08930.2635-0.5315-0.04640.2516-0.06320.11690.0456-0.03610.21820.02370.325667.444785.123321.0616
20.63640.48080.5721-0.1070.35241.1017-0.0488-0.1507-0.08590.002-0.0443-0.05290.0247-0.01380.09540.10930.03480.00160.11520.02590.175950.639782.50518.7922
30.45830.33650.42710.58950.87252.12960.20520.34050.36870.18-0.40260.15721.46140.39270.09361.0560.3980.25260.32530.08020.558748.644764.54380.4905
41.68590.397-0.2240.4777-0.1251.64250.0016-0.226-0.65520.1436-0.0157-0.06850.7474-0.03030.16980.4006-0.06810.00810.09750.10560.226355.894278.70964.8901
50.8331-0.46280.28911.3357-0.47281.3898-0.0409-0.0586-0.07430.15370.01860.05590.1228-0.29760.03050.1714-0.02540.01680.19220.00530.156453.024896.135360.5608
60.0997-0.0262-0.03370.0450.03690.30180.1382-0.203-0.0342-0.02590.1056-0.1122-0.41180.1853-0.13121.045-0.60940.26430.7344-0.45050.548640.516787.066840.1554
70.53020.350.4495-0.03360.39060.8751-0.46950.10450.1691-0.3083-0.13920.1833-0.6328-0.1550.44950.62610.0294-0.00210.3665-0.10350.441164.2663132.274344.7693
80.82160.28310.22710.74090.11431.1128-0.02360.07590.0338-0.07870.0256-0.1272-0.18-0.0074-0.00410.1441-0.02060.03630.1398-0.01480.12160.9729112.107543.3789
90.69310.30052.21182.4875-0.17063.2425-0.1222-0.255-0.37020.680.3002-0.9908-0.0670.2308-0.18540.3091-0.0387-0.23880.3202-0.01010.486976.7272108.425967.2876
10-0.425-0.42080.51240.00570.74891.8691-0.2320.11120.21470.1427-0.37940.2637-0.0979-0.61320.49580.32980.1669-0.02710.9747-0.06690.502113.104389.52461.9175
111.0973-0.21250.50141.5235-0.09241.7191-0.0355-0.00630.0326-0.1377-0.02970.0513-0.0145-0.19530.05450.08990.0417-0.00790.1264-0.01190.068233.397487.29050.2448
120.264-0.152-0.20680.0746-0.03670.1080.07810.2915-0.420.2809-0.0641-0.0590.0959-0.1442-0.02930.69490.3222-0.18590.8958-0.40690.789736.602103.834423.9081
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1(chain A and resid 25:125)A25 - 125
2X-RAY DIFFRACTION2(chain A and resid 126:277)A126 - 277
3X-RAY DIFFRACTION3(chain A and resid 278:299)A278 - 299
4X-RAY DIFFRACTION4(chain B and resid 24:125)B24 - 125
5X-RAY DIFFRACTION5(chain B and resid 126:291)B126 - 291
6X-RAY DIFFRACTION6(chain B and resid 292:299)B292 - 299
7X-RAY DIFFRACTION7(chain C and resid 25:33)C25 - 33
8X-RAY DIFFRACTION8(chain C and resid 34:282)C34 - 282
9X-RAY DIFFRACTION9(chain C and resid 283:299)C283 - 299
10X-RAY DIFFRACTION10(chain D and resid 25:33)D25 - 33
11X-RAY DIFFRACTION11(chain D and resid 34:282)D34 - 282
12X-RAY DIFFRACTION12(chain D and resid 283:299)D283 - 299

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