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- PDB-3mfd: The Structure of the Beta-lactamase superfamily domain of D-alany... -

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Basic information

Entry
Database: PDB / ID: 3mfd
TitleThe Structure of the Beta-lactamase superfamily domain of D-alanyl-D-alanine carboxypeptidase from Bacillus subtilis
ComponentsD-alanyl-D-alanine carboxypeptidase dacB
KeywordsHYDROLASE / penicillin-binding protein 5* / Beta-lactamase / Structural Genomics / PSI-2 / Protein Structure Initiative / Midwest Center for Structural Genomics / MCSG
Function / homology
Function and homology information


serine-type D-Ala-D-Ala carboxypeptidase / serine-type D-Ala-D-Ala carboxypeptidase activity / sporulation resulting in formation of a cellular spore / peptidoglycan biosynthetic process / cell wall organization / regulation of cell shape / endopeptidase activity / membrane
Similarity search - Function
Spermidine Synthase; Chain: A, domain 2 - #30 / Peptidase S11, D-Ala-D-Ala carboxypeptidase A, C-terminal / Penicillin-binding protein 5, C-terminal domain / Penicillin-binding protein 5, C-terminal domain / Peptidase S11, D-alanyl-D-alanine carboxypeptidase A, N-terminal / Peptidase S11, D-alanyl-D-alanine carboxypeptidase A / D-alanyl-D-alanine carboxypeptidase / Spermidine Synthase; Chain: A, domain 2 / Beta-lactamase / DD-peptidase/beta-lactamase superfamily ...Spermidine Synthase; Chain: A, domain 2 - #30 / Peptidase S11, D-Ala-D-Ala carboxypeptidase A, C-terminal / Penicillin-binding protein 5, C-terminal domain / Penicillin-binding protein 5, C-terminal domain / Peptidase S11, D-alanyl-D-alanine carboxypeptidase A, N-terminal / Peptidase S11, D-alanyl-D-alanine carboxypeptidase A / D-alanyl-D-alanine carboxypeptidase / Spermidine Synthase; Chain: A, domain 2 / Beta-lactamase / DD-peptidase/beta-lactamase superfamily / Beta-lactamase/transpeptidase-like / Roll / 3-Layer(aba) Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
CITRIC ACID / D-alanyl-D-alanine carboxypeptidase DacB
Similarity search - Component
Biological speciesBacillus subtilis (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MAD / Resolution: 1.75 Å
AuthorsCuff, M.E. / Rakowski, E. / Buck, K. / Joachimiak, A. / Midwest Center for Structural Genomics (MCSG)
CitationJournal: TO BE PUBLISHED
Title: The Structure of the Beta-lactamase superfamily domain of D-alanyl-D-alanine carboxypeptidase from Bacillus subtilis.
Authors: Cuff, M.E. / Rakowski, E. / Buck, K. / Joachimiak, A.
History
DepositionApr 1, 2010Deposition site: RCSB / Processing site: RCSB
Revision 1.0May 19, 2010Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.2Nov 8, 2017Group: Refinement description / Category: software

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: D-alanyl-D-alanine carboxypeptidase dacB
B: D-alanyl-D-alanine carboxypeptidase dacB
hetero molecules


Theoretical massNumber of molelcules
Total (without water)77,96714
Polymers76,7022
Non-polymers1,26512
Water13,926773
1
A: D-alanyl-D-alanine carboxypeptidase dacB
hetero molecules


Theoretical massNumber of molelcules
Total (without water)39,1758
Polymers38,3511
Non-polymers8257
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: D-alanyl-D-alanine carboxypeptidase dacB
hetero molecules


Theoretical massNumber of molelcules
Total (without water)38,7916
Polymers38,3511
Non-polymers4405
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)100.439, 100.439, 171.853
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number96
Space group name H-MP43212

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Components

#1: Protein D-alanyl-D-alanine carboxypeptidase dacB / DD-carboxypeptidase / DD-peptidase / Penicillin-binding protein 5* / PBP-5*


Mass: 38350.812 Da / Num. of mol.: 2 / Fragment: Beta-lactamase domain residues 27-358
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Bacillus subtilis (bacteria) / Strain: 168 / Gene: BSU23190, dacB, DacC / Plasmid: pMCSG19 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: P35150, serine-type D-Ala-D-Ala carboxypeptidase
#2: Chemical
ChemComp-CIT / CITRIC ACID / Citric acid


Mass: 192.124 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C6H8O7
#3: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C2H6O2
#4: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 773 / 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: 2.83 Å3/Da / Density % sol: 56.46 %
Crystal growTemperature: 277 K / Method: vapor diffusion, sitting drop / pH: 7
Details: 0.2M ammonium citrate tribasic pH 7.0, 20% PEG 3350, VAPOR DIFFUSION, SITTING DROP, temperature 277K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-BM / Wavelength: 0.97948, 0.97923
DetectorType: ADSC QUANTUM 210r / Detector: CCD / Date: Oct 12, 2009
RadiationMonochromator: SAGITALLY FOCUSED Si(111) / Protocol: MAD / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelength
IDWavelength (Å)Relative weight
10.979481
20.979231
ReflectionRedundancy: 15.3 % / Av σ(I) over netI: 35.59 / Number: 1341390 / Rmerge(I) obs: 0.107 / Χ2: 1.9 / D res high: 1.75 Å / D res low: 50 Å / Num. obs: 87904 / % possible obs: 98.6
Diffraction reflection shell
Highest resolution (Å)Lowest resolution (Å)% possible obs (%)IDRmerge(I) obsChi squaredRedundancy
4.755096.810.0855.66114.7
3.774.7599.410.0723.51315.4
3.293.7799.610.0753.02215.7
2.993.2999.910.0932.98516
2.782.9999.910.1062.60816.1
2.612.7810010.1222.32816.2
2.482.6110010.1281.90216.3
2.382.4810010.1431.70316.3
2.282.3810010.161.4916.3
2.22.2810010.1751.34516.3
2.142.210010.1871.21916.3
2.072.1410010.2211.1716.3
2.022.0710010.2561.10216.2
1.972.0210010.3111.02816.2
1.931.9710010.3940.98916.1
1.891.9310010.4771.00115.7
1.851.8910010.5990.9714.1
1.811.8598.210.6770.97812.4
1.781.819210.7940.97711.2
1.751.7885.310.8840.98310.4
ReflectionResolution: 1.75→50 Å / Num. all: 87904 / Num. obs: 87904 / % possible obs: 98.6 % / Observed criterion σ(I): -3 / Redundancy: 15.3 % / Rmerge(I) obs: 0.107 / Net I/σ(I): 9.3
Reflection shellResolution: 1.75→1.78 Å / Redundancy: 10.4 % / Rmerge(I) obs: 0.884 / % possible all: 85.3

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Phasing

PhasingMethod: MAD
Phasing MADD res high: 1.75 Å / D res low: 50 Å / FOM : 0.476 / FOM acentric: 0.494 / FOM centric: 0.302 / Reflection: 86837 / Reflection acentric: 78511 / Reflection centric: 8326
Phasing MAD set

Highest resolution: 1.75 Å / Lowest resolution: 50 Å

IDR cullis acentricR cullis centricLoc acentricLoc centricPower acentricPower centricReflection acentricReflection centric
11.810.80.500785118326
20.90.8521.830.60.770.63620977203
Phasing MAD set shell
IDResolution (Å)R cullis acentricR cullis centricLoc acentricLoc centricPower acentricPower centricReflection acentricReflection centric
111.24-501.5111.8100208132
16.33-11.241.3511.61.1001243416
14.41-6.331.3111.51003144673
13.38-4.411.0811.21.1005942915
12.74-3.381.56110.60095651204
12.31-2.742.1510.90.400140461465
11.99-2.312.1410.60.300195011723
11.75-1.993.0710.50.200248621798
211.24-500.860.9242.447.81.090.82208131
26.33-11.240.820.8234.640.81.240.961243416
24.41-6.330.840.7632.537.31.120.923141672
23.38-4.410.90.8336450.780.595934911
22.74-3.380.890.8425.933.80.810.6295391192
22.31-2.740.90.8419.125.40.810.6139761464
21.99-2.310.940.9216.922.90.630.44194281719
21.75-1.990.960.9616.3210.50.388628698
Phasing MAD set site

Atom type symbol: Se

IDB isoFract xFract yFract zOccupancyOccupancy iso
125.5311-0.937-0.552-0.0595.8060
225.2331-0.907-0.518-0.0045.2690
329.8529-1.025-0.50.0324.8960
427.3116-0.778-0.736-0.135.0750
523.9388-0.722-0.786-0.1754.7650
625.2657-1.08-0.563-0.0395.0910
727.5667-0.999-0.57604.9880
822.8977-0.799-0.71-0.1764.3650
926.3777-0.997-0.521-0.0914.4750
1025.064-0.75-0.668-0.1744.0550
1138.1428-0.883-0.5030.0825.880
1234.4126-0.828-0.717-0.1464.9140
1329.741-0.789-0.653-0.2193.7290
1432.4238-0.741-0.772-0.2353.9460
1526.2973-0.922-0.47-0.0243.3730
1643.786-0.916-0.6160.1234.0020
1757.1226-0.609-0.772-0.2333.830
1847.0811-0.953-0.580.1143.4330
1947.6493-0.488-0.716-0.3272.9040
2047.5528-0.785-0.849-0.0723.1110
2136.9163-0.929-0.7030.0933.3460
2245.1357-1.074-0.695-0.0342.8130
2327.3026-0.962-0.502-0.0412.7460
2449.2945-0.588-0.732-0.3063.2190
2529.1411-0.793-0.851-0.1693.0110
2645.0919-0.986-0.6680.0993.7570
2720.3553-0.937-0.552-0.0593.765-0.12
2822.5666-0.907-0.518-0.0042.995-0.167
2926.2856-1.024-0.50.0322.895-0.17
3022.2103-0.778-0.736-0.133.203-0.164
3119.1377-0.722-0.786-0.1752.908-0.138
3222.3602-1.08-0.563-0.0393.321-0.181
3322.333-0.999-0.57602.827-0.145
3419.4572-0.799-0.71-0.1762.629-0.12
3518.5027-0.997-0.521-0.0912.526-0.094
3620.3046-0.75-0.668-0.1742.706-0.075
3735.1849-0.883-0.5050.0824.231-0.172
3832.762-0.829-0.718-0.1462.714-0.121
3923.8835-0.788-0.653-0.2192.226-0.09
4026.8748-0.741-0.772-0.2352.236-0.113
4124.7231-0.921-0.47-0.0241.738-0.059
4244.1361-0.917-0.6160.1222.35-0.164
4351.2913-0.609-0.772-0.2332.414-0.107
4451.5676-0.953-0.5810.1142.12-0.079
4537.3816-0.488-0.716-0.3271.654-0.098
4645.7938-0.785-0.849-0.0721.939-0.093
4730.5967-0.929-0.7030.0931.763-0.075
4838.9866-1.074-0.695-0.0341.443-0.073
4925.5961-0.962-0.502-0.0411.258-0.049
5040.9943-0.588-0.732-0.3061.965-0.096
5120.6107-0.793-0.851-0.1691.253-0.079
5247.5879-0.985-0.6680.0992.358-0.117
Phasing MAD shell
Resolution (Å)FOM FOM acentricFOM centricReflectionReflection acentricReflection centric
11.24-500.590.7380.356340208132
6.33-11.240.6830.7650.4416591243416
4.41-6.330.6920.7410.46438173144673
3.38-4.410.6370.6740.39368575942915
2.74-3.380.6440.6750.3961076995651204
2.31-2.740.6010.6240.38815511140461465
1.99-2.310.4730.4920.25821224195011723
1.75-1.990.250.2630.06926660248621798
Phasing dmMethod: Solvent flattening and Histogram matching / Reflection: 86837
Phasing dm shell
Resolution (Å)Delta phi finalFOM Reflection
9.49-10053.10.826583
6.71-9.4947.30.9011099
5.48-6.7144.60.9011391
4.74-5.48430.9291624
4.24-4.7441.80.9381824
3.87-4.24450.9361994
3.59-3.8745.30.9322163
3.35-3.59440.932317
3.16-3.3542.30.9222446
3-3.1644.20.9212580
2.86-344.60.9152688
2.74-2.8643.30.9132817
2.63-2.7442.90.9162949
2.54-2.6344.60.9183028
2.45-2.5443.40.9073123
2.37-2.45450.9023244
2.3-2.3747.30.8983347
2.24-2.348.90.8893424
2.18-2.2451.10.8943539
2.12-2.1852.60.8893622
2.07-2.1252.30.8863745
2.02-2.0755.30.8893806
1.98-2.0255.80.8823919
1.94-1.9858.80.8643952
1.9-1.9461.10.8514040
1.86-1.9700.844111
1.83-1.8670.50.8444160
1.79-1.8373.80.8544093
1.75-1.7975.80.825209

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
MLPHAREphasing
DM6phasing
REFMACrefinement
PDB_EXTRACT3.1data extraction
SBC-Collectdata collection
HKL-3000data reduction
HKL-3000data scaling
HKL-3000phasing
SHELXDphasing
SHELXEmodel building
SOLVEphasing
RESOLVEphasing
ARP/wARPmodel building
CCP4phasing
Omodel building
Cootmodel building
RefinementMethod to determine structure: MAD / Resolution: 1.75→34.778 Å / Cor.coef. Fo:Fc: 0.966 / Cor.coef. Fo:Fc free: 0.956 / Occupancy max: 1 / Occupancy min: 0 / SU B: 3.627 / SU ML: 0.053 / SU R Cruickshank DPI: 0.095 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / ESU R: 0.094 / ESU R Free: 0.092
Stereochemistry target values: MAXIMUM LIKELIHOOD WITH PHASES
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.19014 4388 5 %RANDOM
Rwork0.16456 ---
obs0.16583 83341 98.47 %-
all-87729 --
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 15.672 Å2
Baniso -1Baniso -2Baniso -3
1-0.91 Å20 Å20 Å2
2--0.91 Å20 Å2
3----1.82 Å2
Refinement stepCycle: LAST / Resolution: 1.75→34.778 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4958 0 84 773 5815
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0150.0225405
X-RAY DIFFRACTIONr_bond_other_d0.0010.023663
X-RAY DIFFRACTIONr_angle_refined_deg1.3931.9727317
X-RAY DIFFRACTIONr_angle_other_deg0.85239028
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.0755694
X-RAY DIFFRACTIONr_dihedral_angle_2_deg31.94725.064233
X-RAY DIFFRACTIONr_dihedral_angle_3_deg12.82151023
X-RAY DIFFRACTIONr_dihedral_angle_4_deg14.0761521
X-RAY DIFFRACTIONr_chiral_restr0.0880.2811
X-RAY DIFFRACTIONr_gen_planes_refined0.0070.026012
X-RAY DIFFRACTIONr_gen_planes_other0.0020.021028
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it0.751.53281
X-RAY DIFFRACTIONr_mcbond_other0.2321.51340
X-RAY DIFFRACTIONr_mcangle_it1.34325321
X-RAY DIFFRACTIONr_scbond_it2.4132124
X-RAY DIFFRACTIONr_scangle_it3.7424.51979
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
LS refinement shellResolution: 1.75→1.795 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.285 282 -
Rwork0.275 5358 -
obs--86.32 %
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)
11.0834-0.4508-0.59990.81970.5833.10550.07910.0260.0031-0.0262-0.10280.00810.0413-0.19480.02370.0182-0.01340.01560.0313-0.01780.0281-6.632446.552442.0017
22.97390.3571-0.80680.677-0.28390.70240.0301-0.61070.22410.0424-0.102-0.31730.10520.4240.07190.07880.0241-0.02240.3333-0.02160.235222.235449.87357.3807
32.5596-0.92040.51051.1496-0.10041.42170.12240.0213-0.0228-0.0049-0.0385-0.04460.14370.0221-0.08390.08290.041-0.01680.02820.00860.070624.137427.178427.7941
40.99190.31950.08583.0873-0.00441.13380.28870.1492-0.51670.0677-0.1340.39740.6756-0.1488-0.15470.4861-0.0265-0.19330.1079-0.06970.43137.41746.162919.5653
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A-3 - 239
2X-RAY DIFFRACTION2A240 - 331
3X-RAY DIFFRACTION3B-3 - 239
4X-RAY DIFFRACTION4B240 - 331

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