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- PDB-2a81: carboxymethylproline synthase (CarB) from pectobacterium carotovo... -

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Open data


ID or keywords:

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Basic information

Entry
Database: PDB / ID: 2a81
Titlecarboxymethylproline synthase (CarB) from pectobacterium carotovora, complexed with acetyl CoA and bicine
ComponentsCarB
KeywordsBIOSYNTHETIC PROTEIN / carbapenem / carbapenam / crotonase / antibiotic / beta-lactam / bicine / acetyl coenzyme a
Function / homology
Function and homology information


carboxymethylproline synthase / antibiotic biosynthetic process / transferase activity
Similarity search - Function
Single alpha-helices involved in coiled-coils or other helix-helix interfaces - #1610 / Enoyl-CoA hydratase/isomerase / Enoyl-CoA hydratase/isomerase / 2-enoyl-CoA Hydratase; Chain A, domain 1 / 2-enoyl-CoA Hydratase; Chain A, domain 1 / ClpP/crotonase-like domain superfamily / Single alpha-helices involved in coiled-coils or other helix-helix interfaces / Alpha-Beta Complex / Up-down Bundle / Mainly Alpha / Alpha Beta
Similarity search - Domain/homology
ACETYL COENZYME *A / Carboxymethylproline synthase
Similarity search - Component
Biological speciesPectobacterium carotovorum (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.15 Å
AuthorsSleeman, M.C. / Sorensen, J.L. / Batchelar, E.T. / McDonough, M.A. / Schofield, C.J.
CitationJournal: J.Biol.Chem. / Year: 2005
Title: Structural and Mechanistic Studies on Carboxymethylproline Synthase (CarB), a Unique Member of the Crotonase Superfamily Catalyzing the First Step in Carbapenem Biosynthesis.
Authors: Sleeman, M.C. / Sorensen, J.L. / Batchelar, E.T. / McDonough, M.A. / Schofield, C.J.
History
DepositionJul 7, 2005Deposition site: RCSB / Processing site: RCSB
Revision 1.0Aug 23, 2005Provider: repository / Type: Initial release
Revision 1.1Apr 1, 2008Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Derived calculations / Version format compliance
Revision 1.3Oct 11, 2017Group: Refinement description / Category: software
Item: _software.classification / _software.contact_author ..._software.classification / _software.contact_author / _software.contact_author_email / _software.date / _software.language / _software.location / _software.name / _software.type / _software.version
Revision 1.4Aug 23, 2023Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / struct_ncs_dom_lim / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: CarB
B: CarB
C: CarB
hetero molecules


Theoretical massNumber of molelcules
Total (without water)83,7985
Polymers82,8253
Non-polymers9732
Water0
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area6130 Å2
ΔGint-37 kcal/mol
Surface area26080 Å2
MethodPISA, PQS
Unit cell
Length a, b, c (Å)139.605, 139.605, 78.546
Angle α, β, γ (deg.)90.00, 90.00, 120.00
Int Tables number173
Space group name H-MP63
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
31C
41A
51B
61C
71A
81B
91C
101A
111B
121C
131A
141B
151C
161A
171B
181C
191A
201B
211C
221A
231B
241C
251A
261B
271C
281A
291B
301C
311A
321B
331C
341A
351B
361C
371A
381B
391C
401A
411B
421C
431A
441B
451C
461A
471B
481C
491A
501B
511C
521A
531B
541C
551A
561B
571C
581A
591B
601C
611A
621B
631C
641A
651B
661C
671A
681B
691C
701A
711B
721C
731A
741B
751C
761A
771B
781C
791A
801B
811C
821A
831B
841C
851A
861B
871C
881A
891B
901C
911A
921B
931C
941A
951B
961C
971A
981B
991C
12B
22C

NCS domain segments:
Dom-IDComponent-IDEns-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDRefine codeAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
111METMETMETMET1AA11
211METMETMETMET1BB11
311METMETMETMET1CC11
421VALVALPHEPHE5AA2 - 32 - 3
521VALVALPHEPHE5BB2 - 32 - 3
621VALVALPHEPHE5CC2 - 32 - 3
731GLUGLUGLUGLU1AA44
831GLUGLUGLUGLU1BB44
931GLUGLUGLUGLU1CC44
1041GLUGLUGLUGLU5AA55
1141GLUGLUGLUGLU5BB55
1241GLUGLUGLUGLU5CC55
1351ASNASNLEULEU1AA6 - 156 - 15
1451ASNASNLEULEU1BB6 - 156 - 15
1551ASNASNLEULEU1CC6 - 156 - 15
1661ASPASPASPASP5AA1616
1761ASPASPASPASP5BB1616
1861ASPASPASPASP5CC1616
1971HISHISASNASN1AA17 - 1917 - 19
2071HISHISASNASN1BB17 - 1917 - 19
2171HISHISASNASN1CC17 - 1917 - 19
2281LYSLYSLYSLYS5AA2020
2381LYSLYSLYSLYS5BB2020
2481LYSLYSLYSLYS5CC2020
2591HISHISSERSER1AA21 - 2521 - 25
2691HISHISSERSER1BB21 - 2521 - 25
2791HISHISSERSER1CC21 - 2521 - 25
28101ARGARGARGARG5AA2626
29101ARGARGARGARG5BB2626
30101ARGARGARGARG5CC2626
31111THRTHRLEULEU1AA27 - 2827 - 28
32111THRTHRLEULEU1BB27 - 2827 - 28
33111THRTHRLEULEU1CC27 - 2827 - 28
34121GLUGLUGLUGLU4AA2929
35121GLUGLUGLUGLU4BB2929
36121GLUGLUGLUGLU4CC2929
37131THRTHRASPASP1AA30 - 4230 - 42
38131THRTHRASPASP1BB30 - 4230 - 42
39131THRTHRASPASP1CC30 - 4230 - 42
40141ASPASPASPASP6AA4343
41141ASPASPASPASP6BB4343
42141ASPASPASPASP6CC4343
43151SERSERVALVAL1AA44 - 4544 - 45
44151SERSERVALVAL1BB44 - 4544 - 45
45151SERSERVALVAL1CC44 - 4544 - 45
46161ARGARGPHEPHE4AA56 - 5856 - 58
47161ARGARGPHEPHE4BB56 - 5856 - 58
48161ARGARGPHEPHE4CC56 - 5856 - 58
49171ARGARGILEILE6AA72 - 8072 - 80
50171ARGARGILEILE6BB72 - 8072 - 80
51171ARGARGILEILE6CC72 - 8072 - 80
52181ASPASPALAALA1AA81 - 10581 - 105
53181ASPASPALAALA1BB81 - 10581 - 105
54181ASPASPALAALA1CC81 - 10581 - 105
55191ILEILEMETMET4AA106 - 108106 - 108
56191ILEILEMETMET4BB106 - 108106 - 108
57191ILEILEMETMET4CC106 - 108106 - 108
58201GLYGLYMETMET1AA109 - 129109 - 129
59201GLYGLYMETMET1BB109 - 129109 - 129
60201GLYGLYMETMET1CC109 - 129109 - 129
61211PROPROALAALA4AA130 - 142130 - 142
62211PROPROALAALA4BB130 - 142130 - 142
63211PROPROALAALA4CC130 - 142130 - 142
64221ALAALAILEILE1AA143 - 157143 - 157
65221ALAALAILEILE1BB143 - 157143 - 157
66221ALAALAILEILE1CC143 - 157143 - 157
67231ILEILEILEILE2AA158158
68231ILEILEILEILE2BB158158
69231ILEILEILEILE2CC158158
70241TYRTYRGLNGLN1AA159 - 162159 - 162
71241TYRTYRGLNGLN1BB159 - 162159 - 162
72241TYRTYRGLNGLN1CC159 - 162159 - 162
73251SERSERASPASP4AA163 - 165163 - 165
74251SERSERASPASP4BB163 - 165163 - 165
75251SERSERASPASP4CC163 - 165163 - 165
76261ALAALASERSER1AA166 - 197166 - 197
77261ALAALASERSER1BB166 - 197166 - 197
78261ALAALASERSER1CC166 - 197166 - 197
79271TYRTYRILEILE4AA198 - 204198 - 204
80271TYRTYRILEILE4BB198 - 204198 - 204
81271TYRTYRILEILE4CC198 - 204198 - 204
82281ASNASNILEILE1AA205 - 215205 - 215
83281ASNASNILEILE1BB205 - 215205 - 215
84281ASNASNILEILE1CC205 - 215205 - 215
85291HISHISHISHIS3AA216216
86291HISHISHISHIS3BB216216
87291HISHISHISHIS3CC216216
88301LEULEUSERSER5AA217 - 225217 - 225
89301LEULEUSERSER5BB217 - 225217 - 225
90301LEULEUSERSER5CC217 - 225217 - 225
91311ARGARGARGARG3AA4646
92311ARGARGARGARG3BB4646
93311ARGARGARGARG3CC4646
94321ALAALAALAALA1AA47 - 5447 - 54
95321ALAALAALAALA1BB47 - 5447 - 54
96321ALAALAALAALA1CC47 - 5447 - 54
97331GLUGLUGLUGLU5AA5555
98331GLUGLUGLUGLU5BB5555
99331GLUGLUGLUGLU5CC5555
112GLYGLYSERSER4BB62 - 7162 - 71
212GLYGLYSERSER4CC62 - 7162 - 71

NCS ensembles :
ID
1
2
DetailsThe biological unit is a trimer. There is 1 biological unit in the asymmetric unit (chains A, B & C)

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Components

#1: Protein CarB


Mass: 27608.275 Da / Num. of mol.: 3
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Pectobacterium carotovorum (bacteria) / Gene: CarB / Plasmid: pET24a / Species (production host): Escherichia coli / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / References: UniProt: Q9XB60
#2: Chemical ChemComp-ACO / ACETYL COENZYME *A / Acetyl-CoA


Mass: 809.571 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C23H38N7O17P3S
#3: Chemical ChemComp-BCN / BICINE / Bicine


Mass: 163.172 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C6H13NO4 / Comment: pH buffer*YM

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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: 2.7 Å3/Da / Density % sol: 53.6 %
Crystal growTemperature: 291 K / Method: vapor diffusion, hanging drop / pH: 8
Details: PEG 6000, BICINE, pH 8, vapor diffusion, hanging drop, temperature 291K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: SRS / Beamline: PX10.1 / Wavelength: 1.009 Å
DetectorType: MARMOSAIC 225 mm CCD / Detector: CCD / Date: Feb 8, 2005 / Details: mirrors
RadiationMonochromator: Si III / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.009 Å / Relative weight: 1
ReflectionResolution: 3.15→47.91 Å / Num. obs: 15149 / % possible obs: 99.3 % / Redundancy: 5.9 % / Rmerge(I) obs: 0.203 / Χ2: 1.012
Reflection shell
Resolution (Å)% possible obs (%)Redundancy (%)Rmerge(I) obsNum. measured obsΧ2Diffraction-ID
3.15-3.2694.33.90.56214320.9941
3.26-3.3999.15.20.47314900.9991
3.39-3.5599.75.90.43215041.0561
3.55-3.7399.96.20.32115211.0431
3.73-3.971006.20.26115251.0541
3.97-4.271006.20.17714981.0151
4.27-4.71006.20.14915311.0371
4.7-5.381006.20.1515321.0041
5.38-6.781006.20.17915330.9541
6.78-5099.760.06215830.9521

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Phasing

Phasing MR
Highest resolutionLowest resolution
Translation3.05 Å47.669 Å

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
PHASERphasing
REFMAC5.2.0005refinement
PDB_EXTRACT1.7data extraction
HKL-2000data reduction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: monomer of 2A7K
Resolution: 3.15→47.91 Å / Cor.coef. Fo:Fc: 0.922 / Cor.coef. Fo:Fc free: 0.883 / SU B: 44.072 / SU ML: 0.323 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R Free: 0.456 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.232 798 5.3 %RANDOM
Rwork0.192 ---
all0.194 ---
obs0.194 15136 99.23 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso mean: 45.538 Å2
Baniso -1Baniso -2Baniso -3
1-0.75 Å20.38 Å20 Å2
2--0.75 Å20 Å2
3----1.13 Å2
Refinement stepCycle: LAST / Resolution: 3.15→47.91 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms5304 0 52 0 5356
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0090.0225456
X-RAY DIFFRACTIONr_bond_other_d0.0010.024913
X-RAY DIFFRACTIONr_angle_refined_deg1.1141.9477407
X-RAY DIFFRACTIONr_angle_other_deg0.7473.00111321
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.4985694
X-RAY DIFFRACTIONr_dihedral_angle_2_deg42.44824.274248
X-RAY DIFFRACTIONr_dihedral_angle_3_deg16.91315865
X-RAY DIFFRACTIONr_dihedral_angle_4_deg17.2161532
X-RAY DIFFRACTIONr_chiral_restr0.0590.2850
X-RAY DIFFRACTIONr_gen_planes_refined0.0030.026163
X-RAY DIFFRACTIONr_gen_planes_other0.0010.021111
X-RAY DIFFRACTIONr_nbd_refined0.2250.21201
X-RAY DIFFRACTIONr_nbd_other0.170.24969
X-RAY DIFFRACTIONr_nbtor_refined0.180.22735
X-RAY DIFFRACTIONr_nbtor_other0.0870.23209
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1520.2102
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.2560.212
X-RAY DIFFRACTIONr_symmetry_vdw_other0.2160.242
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.2850.24
X-RAY DIFFRACTIONr_mcbond_it0.1821.53553
X-RAY DIFFRACTIONr_mcbond_other0.0421.51415
X-RAY DIFFRACTIONr_mcangle_it0.30725549
X-RAY DIFFRACTIONr_scbond_it0.50532128
X-RAY DIFFRACTIONr_scangle_it0.9034.51858
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION

Ens-IDDom-IDAuth asym-IDNumberRmsTypeWeight
11A22500.02TIGHT POSITIONAL0.05
12B22500.02TIGHT POSITIONAL0.05
13C22500.02TIGHT POSITIONAL0.05
11A5230.44MEDIUM POSITIONAL0.5
12B5230.34MEDIUM POSITIONAL0.5
13C5230.47MEDIUM POSITIONAL0.5
11A2650.96LOOSE POSITIONAL5
12B2650.76LOOSE POSITIONAL5
13C2650.85LOOSE POSITIONAL5
11A22500.03TIGHT THERMAL0.5
12B22500.03TIGHT THERMAL0.5
13C22500.03TIGHT THERMAL0.5
11A5230.28MEDIUM THERMAL2
12B5230.22MEDIUM THERMAL2
13C5230.24MEDIUM THERMAL2
11A2650.95LOOSE THERMAL10
12B2650.76LOOSE THERMAL10
13C2650.77LOOSE THERMAL10
21B1390.31MEDIUM POSITIONAL0.5
21B1390.19MEDIUM THERMAL2
LS refinement shellResolution: 3.15→3.232 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.352 52 -
Rwork0.317 983 -
all-1035 -
obs--93.08 %
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)
12.3030.4190.21421.16960.26582.2421-0.05480.0427-0.0766-0.01470.0750.1083-0.0226-0.4307-0.0202-0.299-0.0376-0.0191-0.07820.0343-0.21289.8797-51.8633-7.3267
22.66630.8046-0.29382.1811-0.82132.679-0.12520.20280.0986-0.28410.1342-0.0059-0.40360.1564-0.009-0.0837-0.0984-0.0391-0.2150.0011-0.227438.224-33.9344-14.5371
30.87150.34650.20012.9444-0.63993.50560.0283-0.0594-0.0620.1167-0.024-0.2257-0.14880.2738-0.0044-0.311-0.0409-0.0666-0.24980.0025-0.197834.4701-48.768116.2067
Refinement TLS group

Refine-ID: X-RAY DIFFRACTION / Selection: ALL

IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11AA1 - 2351 - 235
22BB1 - 2311 - 231
33CC1 - 2311 - 231

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