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- PDB-9b8e: Structure of S-nitrosylated Legionella pneumophila Ceg10. -

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Basic information

Entry
Database: PDB / ID: 9b8e
TitleStructure of S-nitrosylated Legionella pneumophila Ceg10.
ComponentsCeg10
KeywordsHYDROLASE / Bacterial / effector / pathogen
Function / homologyACETATE ION / Type IV secretion protein Dot
Function and homology information
Biological speciesLegionella pneumophila (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.4 Å
AuthorsTomchick, D.R. / Heisler, D.B. / Alto, N.M.
Funding support United States, 6items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)1R35GM137978-01 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)1R21AI169558-01A1 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)R01AI083359 United States
Robert A. Welch FoundationI-1704 United States
Burroughs Wellcome Fund1011019 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)T32 AI007520 United States
CitationJournal: Plos Pathog. / Year: 2024
Title: Exploiting bacterial effector proteins to uncover evolutionarily conserved antiviral host machinery.
Authors: Embry, A. / Baggett, N.S. / Heisler, D.B. / White, A. / de Jong, M.F. / Kocsis, B.L. / Tomchick, D.R. / Alto, N.M. / Gammon, D.B.
History
DepositionMar 29, 2024Deposition site: RCSB / Processing site: RCSB
Revision 1.0Apr 10, 2024Provider: repository / Type: Initial release
Revision 1.1May 29, 2024Group: Database references / Category: citation / citation_author
Item: _citation.country / _citation.journal_abbrev ..._citation.country / _citation.journal_abbrev / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.journal_volume / _citation.page_first / _citation.page_last / _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title / _citation.year / _citation_author.identifier_ORCID / _citation_author.name
Revision 1.2Oct 16, 2024Group: Structure summary / Category: pdbx_entry_details / pdbx_modification_feature / Item: _pdbx_entry_details.has_protein_modification

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Ceg10
B: Ceg10
hetero molecules


Theoretical massNumber of molelcules
Total (without water)54,22512
Polymers53,6752
Non-polymers55010
Water6,215345
1
A: Ceg10
hetero molecules


Theoretical massNumber of molelcules
Total (without water)27,0836
Polymers26,8371
Non-polymers2465
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: Ceg10
hetero molecules


Theoretical massNumber of molelcules
Total (without water)27,1426
Polymers26,8371
Non-polymers3045
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)103.416, 115.836, 40.878
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number18
Space group name H-MP21212
Space group name HallP22ab
Symmetry operation#1: x,y,z
#2: x+1/2,-y+1/2,-z
#3: -x+1/2,y+1/2,-z
#4: -x,-y,z

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Components

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Protein , 1 types, 2 molecules AB

#1: Protein Ceg10


Mass: 26837.357 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Details: Cys159 is nitrosylated. / Source: (gene. exp.) Legionella pneumophila (bacteria) / Gene: C3927_00720 / Plasmid: pET28b / Production host: Escherichia coli (E. coli) / References: UniProt: A0AA44XLE0

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Non-polymers , 5 types, 355 molecules

#2: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: C2H6O2
#3: Chemical ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Mg
#4: Chemical ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Cl
#5: Chemical ChemComp-ACT / ACETATE ION


Mass: 59.044 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C2H3O2
#6: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 345 / Source method: isolated from a natural source / Formula: H2O

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Details

Has ligand of interestY
Has protein modificationY

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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: 2.28 Å3/Da / Density % sol: 46.1 %
Crystal growTemperature: 277 K / Method: vapor diffusion, sitting drop / pH: 7 / Details: 20% PEG 1,000, 0.1 M Tris, 25% ethylene glycol

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-ID / Wavelength: 0.97911 Å
DetectorType: DECTRIS PILATUS3 6M / Detector: PIXEL / Date: Nov 4, 2020
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97911 Å / Relative weight: 1
ReflectionResolution: 1.4→50 Å / Num. obs: 97458 / % possible obs: 99.8 % / Redundancy: 8.9 % / Biso Wilson estimate: 13.24 Å2 / CC1/2: 1 / CC star: 1 / Rmerge(I) obs: 0.058 / Rpim(I) all: 0.02 / Rrim(I) all: 0.061 / Χ2: 0.97 / Net I/σ(I): 8.1
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsNum. unique obsCC1/2CC starRpim(I) allRrim(I) allΧ2% possible all
1.4-1.428.40.88748280.7760.9350.3190.9440.94299.9
1.42-1.458.50.71948170.8320.9530.2580.7650.94499.9
1.45-1.488.60.59948040.8680.9640.2130.6370.94799.9
1.48-1.518.70.46748210.9160.9780.1650.4960.98899.7
1.51-1.548.20.447950.9340.9830.1460.4270.97999.8
1.54-1.588.80.33648360.9580.9890.1180.3570.97999.9
1.58-1.629.30.30148150.9670.9910.1030.319199.9
1.62-1.669.20.25348360.9780.9940.0860.2671.001100
1.66-1.719.10.2148680.9820.9950.0730.2231.002100
1.71-1.768.90.17448040.9870.9970.0610.1851.01699.8
1.76-1.838.30.13648190.990.9970.050.1460.98599.7
1.83-1.99.10.11248460.9940.9980.0390.1191.01699.8
1.9-1.999.40.09148860.9960.9990.0310.0960.991100
1.99-2.099.30.07448570.9970.9990.0250.0780.97199.9
2.09-2.229.10.0649030.9970.9990.0210.0640.97399.8
2.22-2.398.50.05248710.9970.9990.0190.0550.96899.5
2.39-2.639.50.04549260.99810.0150.0470.92100
2.63-3.029.30.0449430.99910.0140.0430.912100
3.02-3.88.70.03549690.99910.0120.0370.94399.3
3.8-508.80.03352140.99910.0110.0350.92599.2

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Processing

Software
NameVersionClassification
PHENIX1.21_5207refinement
HKL-3000data scaling
HKL-3000data reduction
PHENIXphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.4→47.22 Å / SU ML: 0.1426 / Cross valid method: FREE R-VALUE / σ(F): 1.35 / Phase error: 16.9588
Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2
RfactorNum. reflection% reflection
Rfree0.1836 2000 2.11 %
Rwork0.1645 92912 -
obs0.1649 94912 97.21 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.1 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso mean: 20.02 Å2
Refinement stepCycle: LAST / Resolution: 1.4→47.22 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms3639 0 34 345 4018
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00773830
X-RAY DIFFRACTIONf_angle_d0.90665187
X-RAY DIFFRACTIONf_chiral_restr0.0715579
X-RAY DIFFRACTIONf_plane_restr0.0097673
X-RAY DIFFRACTIONf_dihedral_angle_d12.87411423
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.4-1.430.2871040.26884839X-RAY DIFFRACTION71.75
1.43-1.470.241350.22226271X-RAY DIFFRACTION93.15
1.47-1.520.21621430.18986656X-RAY DIFFRACTION98.35
1.52-1.570.22851450.18266733X-RAY DIFFRACTION99.54
1.57-1.620.19991450.16596731X-RAY DIFFRACTION99.94
1.62-1.690.18741460.1696753X-RAY DIFFRACTION99.96
1.69-1.760.191450.1766755X-RAY DIFFRACTION99.81
1.76-1.860.20981460.16426775X-RAY DIFFRACTION99.71
1.86-1.970.16531470.15486806X-RAY DIFFRACTION99.94
1.97-2.130.17811450.14916794X-RAY DIFFRACTION99.94
2.13-2.340.17381470.14526805X-RAY DIFFRACTION99.53
2.34-2.680.16721490.14826893X-RAY DIFFRACTION99.97
2.68-3.370.1691480.16416919X-RAY DIFFRACTION99.52
3.37-47.220.18131550.1687182X-RAY DIFFRACTION99.43
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)
11.23482273781-0.394864911072-0.7730541135651.367354590080.7664202258752.78622114749-0.01149894206010.1763482901010.0302225822425-0.0417885840298-0.01380543720680.05014700970690.0140935666132-0.1687992823930.02625236647360.0474015272977-0.0447778872993-0.001325460697790.07370140945330.01356548769490.0699975330211-19.711464432829.0197329337-7.75790154505
28.032205190443.70539353281-0.05179049514813.242830037940.7893794121552.028333839620.106194362763-0.2378185371020.6367693078640.0732182859119-0.02724602852790.215062108489-0.251372388921-0.00166511253563-0.05641758090250.155694224240.001635766225420.01849926301010.0745843289405-0.0163845696480.106725889709-14.849175513943.568370769910.7361684381
32.914730291520.257026758648-0.7247317165971.302202588530.1914044421681.636160368190.0564656754612-0.0353165541129-0.01455097209970.09076142532330.012261201523-0.0671744154786-0.04917922631180.105954452136-0.05467902057780.067829185658-0.0173516362302-0.008548325260950.07176166955190.0013482444020.0658849906557-8.5838159351335.10686720424.40901826546
43.409063274610.7914422783890.3478382878830.899032678040.3587225259113.029224045220.01145969295740.119974153407-0.00563167217147-0.03786640125090.0750139555995-0.116452518061-0.07983524284770.132431484887-0.06569858632850.0694174553555-0.01845482052440.005769806866270.0692879528551-0.007762732440010.06876067504-8.9487455321233.2019926025-8.03018529216
58.74042586618-0.787396626629-4.262388705347.30457437999-0.2941699392748.668351041330.04185221397890.1236576167980.359114533709-0.1509424520930.0274505306027-0.406834933243-0.3271512628910.194608123166-0.0809005381080.129408081787-0.0266420419806-0.005636715018780.1139156422320.008642615141290.0604184837605-11.431559380635.6510867797-20.452607022
65.72208655480.4740338915013.448312604783.288650471040.04700765767794.24692803841-0.124334873210.4265506416740.609963180427-0.160944425029-0.07164608343120.179768477216-0.4868264754650.01178873465190.213073124060.1507416719030.01955737760960.03635430332940.1256080171730.03282915057670.111065308489-12.62259674942.8715722511-10.8600490245
78.319768876290.8739892219111.297332805484.177082797530.1638149944457.53792538379-0.1709928262460.5817209412280.385588229983-0.2011675895840.1230552266130.0246744024788-0.4582439507980.3346990112560.01107916746440.131301777365-0.1010528115770.006329602138250.2098799259770.02036982026430.1705546485084.216732394143.7298516563-4.01956884092
83.314527649444.190790332760.797017967857.274713753761.334520462341.819081593110.302241255029-0.452437108606-0.5940380771060.4875007018530.01493263294020.2195073764140.318979993817-0.262553971481-0.2935963092730.191583233775-0.0169019763335-0.07039603517360.333556660870.08688757334210.419056055887-0.91762402576828.40742970727.46710521218
93.565310283-0.1332225030123.210387027310.676305391181-0.1305601319224.63952556752-0.150983480174-0.2489898148420.2055701804850.1408006296760.01896819699070.065142995888-0.288210517305-0.4059679350720.1301627798540.1242490690950.01996855937790.01830595744260.1005209362140.01277667221850.0821191349366-28.482642588821.92962760294.02329914124
105.46509715173-0.9875575090943.568681894023.22061387947-1.825320317553.44538742998-0.109613598927-0.01005135120920.03738837519450.14555339388-0.0666444742489-0.2847302467060.04026146977410.2938788728290.1266470141090.07772956528080.000147280947935-0.003053353953360.0981777926234-0.01337568988890.0909167150037-13.540587094915.3644993965-1.01479907707
114.06678430499-3.9850585437-4.039269594886.394383443533.567189747744.088392318540.0332831733227-0.007278664607670.354640447705-0.1923087412940.1094017914440.0524566813799-0.198436190102-0.104936143928-0.1398024841060.1065899281290.00193229552838-0.02344280105930.1159944805110.0006886856531470.0926140259934-32.084109946121.8744260617-10.4117998407
121.51481358867-0.6378458055260.4237354695744.46992890099-0.1236816787871.74742181689-0.0231905618260.141965057611-0.201020279819-0.3052908381230.08290931478340.3536177002650.196398553597-0.192690207489-0.04690283681870.141289354061-0.0426975907863-0.01091237312550.118687600103-0.008383645781420.085038625397-30.04023286127.42459208977-17.1015326444
130.6639488178130.07681787410780.1494699423732.795530857750.0506578329863.17874656653-0.00946557180739-0.032035773054-0.118238517746-0.004135638887940.0952594852714-0.1751585999510.09270043152790.0603691478527-0.03944228235150.0587016978658-0.00706939182792-0.004278334209920.0572325183577-0.01593195602530.0880997614071-21.0548476198.44624826109-2.50348151506
145.39395014411-5.520742776851.188874317958.84827594895-2.21610806478.03129554193-0.0893908458155-0.210086244193-0.4821315554710.1660040828240.09517528124380.1036294296210.4440311291980.006945525033290.00474673365810.141126411888-0.01926094027580.006812266129950.08498774728130.01144790454630.104311749259-24.96838749479.4241863663812.6569173143
153.23858342575-1.26633890636-1.550688042945.414524582913.631339761514.74764921002-0.16715308521-0.152843642353-0.2524757051240.18178224315-0.09879981240210.5271110010880.137368149657-0.4486066207010.2621345784710.0909160151851-0.0128473564688-0.000286075717140.1355352818730.02223237836480.104778474163-31.69339656936.290215235892.81075895533
161.91648081934-1.763465897-0.7387113689897.27193889076-0.08439503100156.48506344632-0.23840343621-0.0708098500284-0.229355792790.5763179418930.1890048993790.3203601430950.310974097667-0.3569602840220.02736185725240.278377948826-0.0194844049079-0.01454818385770.108188092710.006424730814460.243708222375-25.884660344-7.07419840974-1.44776405977
173.441303710233.73275970793-2.173149257418.94703384506-4.911773673084.821635298-0.0760745513540.326827817195-0.230208111786-0.774072574280.302506173513-0.751099194126-0.1255540826750.261419674026-0.2102934251180.344136371589-0.0416941816680.09399979935140.180321615884-0.1124869298050.374356223789-18.1089827855-5.82186090741-13.8976685914
182.731469302481.66252863371-0.2068224155457.665389261464.690448862254.20715349011-0.05346458742390.695989990452-0.2406962435830.09044492148840.589941360423-1.252865402010.1409183179220.357091477534-0.4817528125170.4467109560740.03620200166430.1871122774850.356959503014-0.02935273587840.433439538547-11.07357201329.44860235091-14.8774911178
Refinement TLS group

Refine-ID: X-RAY DIFFRACTION

IDRefine TLS-IDSelection detailsAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11chain 'A' and (resid 57 through 115 )AA57 - 1151 - 59
22chain 'A' and (resid 116 through 139 )AA116 - 13960 - 83
33chain 'A' and (resid 140 through 175 )AA140 - 17584 - 119
44chain 'A' and (resid 176 through 212 )AA176 - 212120 - 156
55chain 'A' and (resid 213 through 225 )AA213 - 225157 - 169
66chain 'A' and (resid 226 through 238 )AA226 - 238170 - 182
77chain 'A' and (resid 239 through 264 )AA239 - 264183 - 202
88chain 'A' and (resid 265 through 284 )AA265 - 284203 - 222
99chain 'B' and (resid 57 through 86 )BG57 - 861 - 30
1010chain 'B' and (resid 87 through 105 )BG87 - 10531 - 49
1111chain 'B' and (resid 106 through 115 )BG106 - 11550 - 59
1212chain 'B' and (resid 116 through 158 )BG116 - 15860 - 102
1313chain 'B' and (resid 159 through 212 )BG159 - 212103 - 156
1414chain 'B' and (resid 213 through 225 )BG213 - 225157 - 169
1515chain 'B' and (resid 226 through 238 )BG226 - 238170 - 182
1616chain 'B' and (resid 239 through 258 )BG239 - 258183 - 199
1717chain 'B' and (resid 259 through 272 )BG259 - 272200 - 213
1818chain 'B' and (resid 273 through 284 )BG273 - 284214 - 225

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