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- PDB-8d1x: Crystal Structure of aminopeptidase A from Neisseria gonorrhoeae -

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Basic information

Entry
Database: PDB / ID: 8d1x
TitleCrystal Structure of aminopeptidase A from Neisseria gonorrhoeae
ComponentsProbable cytosol aminopeptidase
KeywordsHYDROLASE / SSGCID / aminopeptidase A / Neisseria gonorrhoeae / Structural Genomics / Seattle Structural Genomics Center for Infectious Disease
Function / homology
Function and homology information


bacterial leucyl aminopeptidase / leucyl aminopeptidase / metalloaminopeptidase activity / manganese ion binding / cytoplasm
Similarity search - Function
Peptidase M17, leucyl aminopeptidase, N-terminal / Cytosol aminopeptidase family, N-terminal domain / Peptidase M17, leucine aminopeptidase / Cytosol aminopeptidase signature. / Peptidase M17, leucyl aminopeptidase, C-terminal / Peptidase M17, leucine aminopeptidase/peptidase B / Cytosol aminopeptidase family, catalytic domain / Macro domain-like
Similarity search - Domain/homology
ACETATE ION / : / D(-)-TARTARIC ACID / Probable cytosol aminopeptidase
Similarity search - Component
Biological speciesNeisseria gonorrhoeae NCCP11945 (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.8 Å
AuthorsSeattle Structural Genomics Center for Infectious Disease / Seattle Structural Genomics Center for Infectious Disease (SSGCID)
Funding support United States, 1items
OrganizationGrant numberCountry
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)HHSN272201700059C United States
CitationJournal: to be published
Title: Crystal Structure of aminopeptidase A from Neisseria gonorrhoeae
Authors: Abendroth, J. / DeBouver, N.D. / Lorimer, D.D. / Horanyi, P.S. / Edwards, T.E.
History
DepositionMay 27, 2022Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jun 8, 2022Provider: repository / Type: Initial release
Revision 1.1Oct 18, 2023Group: Data collection / Refinement description
Category: chem_comp_atom / chem_comp_bond / pdbx_initial_refinement_model

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Probable cytosol aminopeptidase
B: Probable cytosol aminopeptidase
C: Probable cytosol aminopeptidase
D: Probable cytosol aminopeptidase
E: Probable cytosol aminopeptidase
F: Probable cytosol aminopeptidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)308,33728
Polymers306,6586
Non-polymers1,67922
Water7,350408
1


  • Idetical with deposited unit
  • defined by author
  • Evidence: gel filtration
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)96.610, 93.250, 179.470
Angle α, β, γ (deg.)90.000, 101.324, 90.000
Int Tables number4
Space group name H-MP1211
Space group name HallP2yb
Symmetry operation#1: x,y,z
#2: -x,y+1/2,-z
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
d_1ens_1(chain "A" and ((resid 2 and (name N or name...
d_2ens_1(chain "B" and (resid 2 through 5 or (resid 6...
d_3ens_1(chain "C" and (resid 2 through 44 or (resid 45...
d_4ens_1(chain "D" and ((resid 2 and (name N or name...
d_5ens_1(chain "E" and (resid 2 through 8 or (resid 9...
d_6ens_1(chain "F" and ((resid 2 and (name N or name...

NCS domain segments:
Dom-IDComponent-IDEns-IDBeg label comp-IDEnd label comp-IDLabel asym-IDLabel seq-ID
d_11ens_1GLULEUA2 - 468
d_12ens_1MNMNB
d_13ens_1CLCLC
d_14ens_1TARTARD
d_21ens_1GLULEUF2 - 468
d_22ens_1MNMNG
d_23ens_1CLCLH
d_24ens_1TARTARI
d_31ens_1GLULEUK1 - 467
d_32ens_1MNMNL
d_33ens_1CLCLM
d_34ens_1TARTARN
d_41ens_1GLULEUO3 - 469
d_42ens_1MNMNP
d_43ens_1CLCLQ
d_44ens_1TARTARR
d_51ens_1GLULEUS2 - 468
d_52ens_1MNMNT
d_53ens_1CLCLU
d_54ens_1TARTARV
d_61ens_1GLULEUX3 - 469
d_62ens_1MNMNY
d_63ens_1CLCLZ
d_64ens_1TARTARAA

NCS oper:
IDCodeMatrixVector
1given(0.319910664122, 0.681252626283, -0.658446676781), (0.681437013485, -0.648286191278, -0.339659551391), (-0.658255849677, -0.340029224304, -0.671624421076)19.0089391026, 12.2127537664, 50.5390126903
2given(-0.0687647969063, 0.599281139256, 0.797579788384), (0.599069353461, -0.614478709282, 0.513353509371), (0.797738874852, 0.513106257974, -0.316756460988)-54.3536544894, -25.9301945754, 83.0005354728
3given(-0.14652535741, -0.705471303578, -0.693426679227), (-0.568204575048, 0.633836435377, -0.52478084386), (0.809736920568, 0.317114510889, -0.493725132491)-8.14561258114, 3.56281508579, 88.5495279101
4given(-0.141306239008, -0.565757153213, 0.812373922776), (-0.701935875971, 0.63591759686, 0.320772249469), (-0.698082067412, -0.524907280949, -0.486984366858)-70.8762167707, -36.0963134246, 39.5601922
5given(-0.96651085562, -0.00170563927684, -0.256620063058), (-0.0110432425374, -0.998775173333, 0.0482306948666), (-0.256388012129, 0.049449407759, 0.965308211562)-39.428155481, -26.0823079854, -4.47583507834

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Components

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Protein , 1 types, 6 molecules ABCDEF

#1: Protein
Probable cytosol aminopeptidase / Leucine aminopeptidase / LAP / Leucyl aminopeptidase


Mass: 51109.688 Da / Num. of mol.: 6 / Fragment: NegoA.00799.a.B1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Neisseria gonorrhoeae NCCP11945 (bacteria)
Strain: NCCP11945 / Gene: pepA, NGK_0283 / Plasmid: NegoA.00799.a.B1 / Production host: Escherichia coli BL21(DE3) (bacteria)
References: UniProt: B4RJ22, leucyl aminopeptidase, bacterial leucyl aminopeptidase

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Non-polymers , 5 types, 430 molecules

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


Mass: 54.938 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: Mn / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical
ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: Cl
#4: Chemical
ChemComp-TAR / D(-)-TARTARIC ACID


Mass: 150.087 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: C4H6O6
#5: Chemical
ChemComp-ACT / ACETATE ION


Mass: 59.044 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C2H3O2
#6: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 408 / 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: 2.59 Å3/Da / Density % sol: 52.58 %
Crystal growTemperature: 287 K / Method: vapor diffusion, sitting drop / pH: 8.5
Details: Molecular Dimensions / Calibre Morpheus screen, condition G6: 10% w/v PEG 8000, 20% v/v ethylene glycol: 20mM of each sodium formate, ammonium acetate, trisodium citrate, sodium potassium L- ...Details: Molecular Dimensions / Calibre Morpheus screen, condition G6: 10% w/v PEG 8000, 20% v/v ethylene glycol: 20mM of each sodium formate, ammonium acetate, trisodium citrate, sodium potassium L-tartrate, sodium oxamate, 100mM MOPS/HEPES-Na pH 7.5: NegoA.00799.a.B1.PW37906 at 19mg/ml + 2mM MnCl2: tray 273710 g6: cryo: direct: puck ifb3-2.

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 21-ID-F / Wavelength: 0.97872 Å
DetectorType: RAYONIX MX-300 / Detector: CCD / Date: Aug 4, 2016 / Details: Beryllium Lenses
RadiationMonochromator: Diamond [111] / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97872 Å / Relative weight: 1
ReflectionResolution: 2.8→50 Å / Num. obs: 77236 / % possible obs: 99.9 % / Redundancy: 4.263 % / Biso Wilson estimate: 43.878 Å2 / CC1/2: 0.992 / Rmerge(I) obs: 0.122 / Rrim(I) all: 0.139 / Χ2: 0.968 / Net I/σ(I): 9.61
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique obsCC1/2Rrim(I) all% possible all
2.8-2.874.3020.5512.5856670.8270.63100
2.87-2.954.2930.453.1155430.870.51599.9
2.95-3.044.2930.3683.8653880.90.421100
3.04-3.134.290.3214.3652530.9230.36799.9
3.13-3.234.2960.2565.350790.9490.29399.9
3.23-3.354.2890.2166.2749110.9610.247100
3.35-3.474.2910.1687.8547500.9750.19299.9
3.47-3.614.2850.1468.9445620.9790.16799.9
3.61-3.784.2810.12210.3743510.9850.139100
3.78-3.964.2750.10311.8342110.9880.118100
3.96-4.174.260.0913.3339990.990.103100
4.17-4.434.2640.08414.5337760.990.09699.8
4.43-4.734.2570.07715.7235420.9920.08899.9
4.73-5.114.2460.07815.4933210.9930.08999.8
5.11-5.64.2480.08114.9830540.9910.09299.7
5.6-6.264.2290.08214.9127620.9910.09499.8
6.26-7.234.1940.07216.3524680.9920.08299.7
7.23-8.854.150.06119.4420800.9940.07100
8.85-12.524.0860.05621.1716220.9930.06599.7
12.52-503.7510.05520.428970.9930.06496.1

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Processing

Software
NameVersionClassification
XDSdata reduction
XSCALEdata scaling
PHENIX1.20.1refinement
PDB_EXTRACT3.27data extraction
MoRDaphasing
PHENIXmodel building
Cootmodel building
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: pdb entry 3jru as per Morda
Resolution: 2.8→48.17 Å / SU ML: 0.3028 / Cross valid method: FREE R-VALUE / σ(F): 1.35 / Phase error: 21.1427
Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2
RfactorNum. reflection% reflectionSelection details
Rfree0.2025 2015 2.61 %0
Rwork0.1698 75202 --
obs0.1706 77217 99.87 %-
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso mean: 48.52 Å2
Refinement stepCycle: LAST / Resolution: 2.8→48.17 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms20666 0 88 408 21162
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.004221137
X-RAY DIFFRACTIONf_angle_d0.626928788
X-RAY DIFFRACTIONf_chiral_restr0.04433333
X-RAY DIFFRACTIONf_plane_restr0.00453791
X-RAY DIFFRACTIONf_dihedral_angle_d12.48847298
Refine LS restraints NCS
Ens-IDDom-IDAuth asym-IDRefine-IDTypeRms dev position (Å)
ens_1d_2AX-RAY DIFFRACTIONTorsion NCS0.290943133448
ens_1d_3AX-RAY DIFFRACTIONTorsion NCS0.346020516024
ens_1d_4AX-RAY DIFFRACTIONTorsion NCS0.298851520776
ens_1d_5AX-RAY DIFFRACTIONTorsion NCS0.308170258732
ens_1d_6AX-RAY DIFFRACTIONTorsion NCS0.311163940848
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.8-2.870.28871420.24685313X-RAY DIFFRACTION99.98
2.87-2.950.28481180.23995381X-RAY DIFFRACTION99.96
2.95-3.030.27591310.23165371X-RAY DIFFRACTION99.95
3.03-3.130.28511420.23675347X-RAY DIFFRACTION99.95
3.13-3.240.29481540.23425300X-RAY DIFFRACTION99.96
3.24-3.370.26041640.21565360X-RAY DIFFRACTION100
3.37-3.530.23221550.19745333X-RAY DIFFRACTION99.87
3.53-3.710.23951350.18615376X-RAY DIFFRACTION99.93
3.71-3.950.191270.15615372X-RAY DIFFRACTION99.98
3.95-4.250.16591360.13635399X-RAY DIFFRACTION99.95
4.25-4.680.13711720.11935346X-RAY DIFFRACTION99.87
4.68-5.350.16941230.13155426X-RAY DIFFRACTION99.82
5.35-6.740.2121340.15485419X-RAY DIFFRACTION99.78
6.74-48.170.14481820.13415459X-RAY DIFFRACTION99.26
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)
13.38601977718-0.2100986508741.113214293521.75011667999-0.7980906288623.678022849680.1054523211520.093970261225-0.0853407064702-0.198949068792-0.138057873348-0.0511567202603-0.2997913858090.3265986206220.02761511601950.404730499464-0.04935647471030.121512080020.285286419719-0.01644851936860.363785619825-6.23740064991-10.8385058747-6.85717191435
20.903317433302-0.1249685819880.3437146489760.817861691163-0.3621641211952.70159194693-0.002693102491890.126599141534-0.1907352953430.01467987999710.027395034534-0.261036203110.1319030198550.237715222017-0.04748800567020.290713502799-0.02004610752710.01668786117150.224303254707-0.04473316128220.4704932629912.44726108331-25.805559429218.1051357862
30.341237519248-0.3596202723890.4183903018460.9498797201860.44825192792.30244630540.0338249365197-0.0446069034314-0.144179743159-0.02362207668390.0394204773777-0.000518004704145-0.1007997201020.172461067131-0.08293324040430.25978446506-0.1066333514630.01089868603490.2796098103190.04219209517260.4419543807482.92518567764-15.513468388832.3519178567
40.856522956529-0.1974784366330.7181810719310.92465335168-0.2349653528431.79624678401-0.0863773824936-0.09413429246070.2894368873590.0920883289425-0.11914255307-0.306263392144-0.3666334722070.4131559795180.2464204150580.472136347076-0.150851032905-0.04071108757460.38920487094-0.0009879789184730.4710460603275.4092232286720.512690280257.214214389
51.2877142651-0.01563144654360.7897351232590.6773855401090.709381078751.33531998325-0.0342837922199-0.1552509184030.104790306262-0.0521230367161-0.057414315523-0.0239060212658-0.238052287243-0.06643009278070.0793270810040.403128018254-0.0834828840930.03484010197230.213850412540.03085712755930.325047335258-13.58151805315.156332025132.1421538711
60.43760734740.592952250392-0.2428281078430.930879674439-0.3900499436340.173873942540.0840945531558-0.545166238259-0.01571470688970.267244562091-0.09067190075050.2867401032650.316851722058-0.653120007426-0.07662980750730.624959247501-0.3342707798420.1784389808161.449000077240.08110540453040.579862992518-63.7654128493-19.698403090872.7399364615
71.507807235290.4954224425040.06303307928711.2111505617-0.5045458506050.853551698664-0.0287307261375-0.389665893540.1116595095550.152861527701-0.03486922376720.19434089644-0.1231441619-0.5292386884030.05327659483570.35985322162-0.0007686167850910.04964147357160.649605544782-0.09834611492350.314809923203-40.2450452174.3876078504965.929337636
81.060774968560.8645455283670.2164834876372.033681152260.3463561181750.7958673075580.216090189915-0.3866211250380.03128535002750.559710404727-0.201686504307-0.259830351164-0.08254034051240.0830764923521-0.02207133515490.513832727561-0.12755173475-0.1005570320910.5220819352120.05185368875910.3583158408413.18217739413-6.2137746643179.8837516025
91.359378467650.4363955394620.3189106032240.9797068213450.4529445737341.128772217270.221844723426-0.364753549619-0.2397844275440.156571090342-0.0548556904075-0.02331158097950.271779511261-0.232352985395-0.1669668547270.452115920617-0.173300851917-0.07062613848990.4829940051720.1542343100910.361384196587-18.8325095917-27.038010099569.6594055244
100.152996152978-0.2101531858990.07038986396330.3606398528590.1617245106270.8446345991440.116395607637-0.321261490443-0.1984012974050.203064988175-0.08530652135210.3791418619330.250729805886-0.81967909418-0.2297656263030.531181432697-0.596057755061-0.03943236461661.099779214090.2951785789550.693881858629-62.998253781-43.274864881950.614612164
111.308440401950.4582377675330.06909651892410.737767440309-0.1239047842330.8234899232610.246742062197-0.173540577258-0.1461506352940.117257601273-0.0627437152231-0.0993503872080.279828892729-0.158370909063-0.1362362419410.444803351318-0.146858701802-0.1141343728370.2869802611260.1217743724260.429722061256-34.5473565834-40.307823416631.3891857396
124.604255954050.413257001167-0.5258743512650.2076336029140.5981329021513.034165825530.173586554020.144753491573-0.172686226716-0.142990215562-0.07731628487280.05468057052160.02141094507360.0379072245443-0.09352507800150.4897949030280.0586256638898-0.05361974926420.19550386147-0.00259857365170.392990198868-31.7366346593-15.6058468-9.74646355696
131.10849150749-0.1153888272550.9858924668871.142772495031.688217668853.702223330640.05831394365170.110649202953-0.173769142212-0.3289337686010.0262108716330.045992583736-0.2191219707540.0855932213594-0.09217517519820.3957202681190.0151606370686-0.05191692573510.1683470808480.01896538004630.367878409315-40.5418819834-5.65553594450.339359419771
140.3806992007560.7422231370410.01325128822771.764502828230.2162670225262.476228250120.00375280592088-0.1907742199870.2530706244420.00836151892943-0.09182498532320.274818704195-0.425478628222-0.3749815219750.1263090498920.3510457041310.08918019904980.001560386220550.3294256031360.006921630898580.408498938523-53.87616929435.0766324096121.8415000966
150.336912278719-0.326916254397-0.02184514783020.8545593835750.1794296941122.64864054806-0.00874985221974-0.2202186251470.0235727012670.0215725479393-0.02281629158880.05532782133840.045904637683-0.2341415153670.0181076083880.239425973667-0.06649645037630.004402429413530.3354003011670.03656136458140.361948244886-50.050255168-9.5149468968425.7915888936
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 98 )AA1 - 981 - 98
22chain 'A' and (resid 99 through 227 )AA99 - 22799 - 227
33chain 'A' and (resid 228 through 468 )AA228 - 468228 - 468
44chain 'B' and (resid 1 through 189 )BF1 - 1891 - 189
55chain 'B' and (resid 190 through 468 )BF190 - 468190 - 468
66chain 'C' and (resid 2 through 167 )CK2 - 1671 - 166
77chain 'C' and (resid 168 through 468 )CK168 - 468167 - 467
88chain 'D' and (resid 0 through 189 )DO0 - 1891 - 190
99chain 'D' and (resid 190 through 468 )DO190 - 468191 - 469
1010chain 'E' and (resid 1 through 167 )ES1 - 1671 - 167
1111chain 'E' and (resid 168 through 468 )ES168 - 468168 - 468
1212chain 'F' and (resid 0 through 98 )FX0 - 981 - 99
1313chain 'F' and (resid 99 through 167 )FX99 - 167100 - 168
1414chain 'F' and (resid 168 through 247 )FX168 - 247169 - 248
1515chain 'F' and (resid 248 through 468 )FX248 - 468249 - 469

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