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- PDB-5ht0: Crystal structure of an Antibiotic_NAT family aminoglycoside acet... -

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Basic information

Entry
Database: PDB / ID: 5ht0
TitleCrystal structure of an Antibiotic_NAT family aminoglycoside acetyltransferase HMB0038 from an uncultured soil metagenomic sample in complex with coenzyme A
ComponentsAminoglycoside acetyltransferase HMB0005
KeywordsTRANSFERASE / Antibiotic_NAT family / ACETYLTRANSFERASE / AMINOGLYCOSIDE / ANTIBIOTIC RESISTANCE / METAGENOME / SOIL / COENZYME A / CSGID / Center for Structural Genomics of Infectious Diseases
Function / homologyaminoglycoside 3-N-acetyltransferase activity / Aminoglycoside N(3)-acetyltransferase / Aminoglycoside 3-N-acetyltransferase / Aminoglycoside 3-N-acetyltransferase-like / Transferases; Acyltransferases; Transferring groups other than aminoacyl groups / response to antibiotic / COENZYME A / Aminoglycoside N(3)-acetyltransferase
Function and homology information
Biological speciesuncultured bacterium (environmental samples)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.752 Å
AuthorsXu, Z. / Stogios, P.J. / Wawrzak, Z. / Skarina, T. / Yim, V. / Savchenko, A. / Anderson, W.F. / Center for Structural Genomics of Infectious Diseases (CSGID)
CitationJournal: Commun Biol / Year: 2022
Title: Structural and molecular rationale for the diversification of resistance mediated by the Antibiotic_NAT family.
Authors: Stogios, P.J. / Bordeleau, E. / Xu, Z. / Skarina, T. / Evdokimova, E. / Chou, S. / Diorio-Toth, L. / D'Souza, A.W. / Patel, S. / Dantas, G. / Wright, G.D. / Savchenko, A.
History
DepositionJan 26, 2016Deposition site: RCSB / Processing site: RCSB
Revision 1.0Mar 2, 2016Provider: repository / Type: Initial release
Revision 1.1Apr 6, 2022Group: Database references / Derived calculations
Category: citation / citation_author ...citation / citation_author / database_2 / pdbx_struct_oper_list
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 / _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_struct_oper_list.symmetry_operation
Revision 1.2Sep 27, 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

Downloads & links

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Assembly

Deposited unit
A: Aminoglycoside acetyltransferase HMB0005
B: Aminoglycoside acetyltransferase HMB0005
C: Aminoglycoside acetyltransferase HMB0005
D: Aminoglycoside acetyltransferase HMB0005
E: Aminoglycoside acetyltransferase HMB0005
F: Aminoglycoside acetyltransferase HMB0005
hetero molecules


Theoretical massNumber of molelcules
Total (without water)179,14333
Polymers172,5216
Non-polymers6,62327
Water6,125340
1
A: Aminoglycoside acetyltransferase HMB0005
B: Aminoglycoside acetyltransferase HMB0005
hetero molecules


Theoretical massNumber of molelcules
Total (without water)59,90713
Polymers57,5072
Non-polymers2,40011
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
C: Aminoglycoside acetyltransferase HMB0005
D: Aminoglycoside acetyltransferase HMB0005
hetero molecules


Theoretical massNumber of molelcules
Total (without water)59,61810
Polymers57,5072
Non-polymers2,1118
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
3
E: Aminoglycoside acetyltransferase HMB0005
F: Aminoglycoside acetyltransferase HMB0005
hetero molecules


Theoretical massNumber of molelcules
Total (without water)59,61810
Polymers57,5072
Non-polymers2,1118
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)107.016, 159.501, 146.225
Angle α, β, γ (deg.)90.00, 94.74, 90.00
Int Tables number5
Space group name H-MC121
Components on special symmetry positions
IDModelComponents
11A-305-

SO4

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Components

#1: Protein
Aminoglycoside acetyltransferase HMB0005


Mass: 28753.482 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) uncultured bacterium (environmental samples)
Plasmid: pMCSG53 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: A0A059X981
#2: Chemical
ChemComp-COA / COENZYME A


Mass: 767.534 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: C21H36N7O16P3S
#3: Chemical...
ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 21 / Source method: obtained synthetically / Formula: SO4
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 340 / 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: 3.61 Å3/Da / Density % sol: 65.91 %
Crystal growTemperature: 298 K / Method: vapor diffusion, sitting drop / pH: 11
Details: 2 M ammonium sulfate, 0.5 M Capso pH 11, 10mM sisomycin, 5mM coenzyme A

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 21-ID-G / Wavelength: 0.97856 Å
DetectorType: MARMOSAIC 300 mm CCD / Detector: CCD / Date: Oct 31, 2015
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97856 Å / Relative weight: 1
ReflectionResolution: 2.75→25 Å / % possible obs: 96.7 % / Redundancy: 4 % / Rmerge(I) obs: 0.074 / Net I/σ(I): 17.76
Reflection shellResolution: 2.75→2.8 Å / Redundancy: 3.7 % / Rmerge(I) obs: 0.44 / Mean I/σ(I) obs: 3.19 / % possible all: 90.4

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Processing

Software
NameVersionClassification
PHENIX1.9_1692refinement
HKL-3000data reduction
HKL-3000data scaling
BALBESphasing
PHENIXmodel building
Cootmodel building
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 2NYG
Resolution: 2.752→24.973 Å / SU ML: 0.34 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 25.33 / Stereochemistry target values: ML
RfactorNum. reflection% reflectionSelection details
Rfree0.2331 2021 3.36 %Random selection
Rwork0.2038 ---
obs0.2048 60061 95.13 %-
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.752→24.973 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms12016 0 393 340 12749
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.01412694
X-RAY DIFFRACTIONf_angle_d1.82717304
X-RAY DIFFRACTIONf_dihedral_angle_d16.9674646
X-RAY DIFFRACTIONf_chiral_restr0.0661830
X-RAY DIFFRACTIONf_plane_restr0.0062228
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.7525-2.82120.30771210.31283197X-RAY DIFFRACTION74
2.8212-2.89740.30571290.314022X-RAY DIFFRACTION92
2.8974-2.98250.3391530.29243977X-RAY DIFFRACTION92
2.9825-3.07860.38721240.29764023X-RAY DIFFRACTION93
3.0786-3.18850.27141420.27574092X-RAY DIFFRACTION93
3.1885-3.31580.27331530.24584084X-RAY DIFFRACTION95
3.3158-3.46640.25271410.22524199X-RAY DIFFRACTION97
3.4664-3.64860.26491460.21114273X-RAY DIFFRACTION98
3.6486-3.87640.20131450.18364352X-RAY DIFFRACTION100
3.8764-4.17440.19371620.1714365X-RAY DIFFRACTION100
4.1744-4.59220.18971470.15414358X-RAY DIFFRACTION100
4.5922-5.25130.17611510.1544363X-RAY DIFFRACTION100
5.2513-6.59590.21891510.18914368X-RAY DIFFRACTION100
6.5959-24.97430.21921560.18254367X-RAY DIFFRACTION98
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)
16.7709-0.83470.57284.00820.32493.8448-0.068-0.18470.0009-0.00430.1353-0.5161-0.03620.5719-0.03570.25140.01040.04820.21450.02490.2708117.5248-14.9431306.5516
23.1793-2.6058-4.56252.14123.73816.5493-0.8014-1.0749-1.95752.32890.03711.5551.3383-0.54320.64621.5514-0.10540.54150.83750.02721.389386.2763-5.1955310.6624
31.241-0.26890.71353.3624-0.23071.40660.03790.03780.0689-0.03430.06650.2840.0115-0.0769-0.09610.1810.0432-0.01330.26180.00860.2635100.5065-16.0418302.1898
45.23643.2387-3.67726.8742-6.16656.64380.07340.27550.1546-0.02221.00981.51380.3119-0.724-1.02970.2896-0.0136-0.0230.3643-0.00020.815886.329-11.4739299.5727
52.4462-1.3930.81584.0325-1.54461.62910.0581-0.5017-0.2180.19270.06750.46250.2306-0.1899-0.16150.3109-0.01810.02580.3283-0.01690.2919100.217-27.411311.0479
62.38721.68772.13322.21480.86432.30870.2675-0.6960.25760.25310.16850.5845-0.02-0.8668-0.43350.381-0.00660.04840.4661-0.0090.277489.413211.9303308.2432
77.9626-0.2602-0.3832.72020.05173.7379-0.00170.0155-0.005-0.05080.0478-0.0730.0169-0.0547-0.10860.20750.0109-0.01440.1283-0.0440.2402102.421112.7973304.4606
83.7086-4.955-1.79047.5023-0.29689.1376-0.2736-0.30382.64172.4105-0.69720.4749-0.4455-0.21481.01171.2231-0.05-0.37441.1231-0.49271.9664130.64392.8153315.1645
92.6973-1.0879-0.63462.30480.49170.50220.004-0.20790.32130.11510.1414-0.4017-0.05910.2907-0.13510.2724-0.0364-0.03660.3239-0.04010.3343120.566213.5876304.4707
109.09891.5778-0.59855.3267-0.44354.2086-0.0835-0.50850.81230.19320.1323-0.1662-0.4630.36440.02660.31480.02390.03820.2634-0.08530.2898110.535629.0673313.779
117.93291.1782-0.47958.9491-6.20927.3639-0.2009-0.16780.26040.6104-0.1314-1.093-0.75680.74880.28940.4412-0.0456-0.11640.5086-0.12270.5163121.918852.3893332.4154
122.03931.6144-0.31573.04140.81272.3817-0.29270.68820.56960.12410.11380.03460.2060.29530.32030.41520.0916-0.10490.49470.01620.3282113.927140.8537331.7493
133.1560.8897-3.85538.3568-5.87798.7646-0.6116-1.4902-0.23340.27470.7650.93440.37560.3255-0.29730.54870.0616-0.00080.6045-0.0370.374890.923936.4819330.6356
143.06790.6230.92512.73-0.75412.81080.0488-0.5359-0.30840.4195-0.0467-0.19970.1343-0.23660.01130.4546-0.0026-0.02870.47070.01030.2953107.718334.797337.6365
155.21052.3044-1.63916.6232-3.34177.4270.2207-1.1994-0.15111.2856-0.2235-0.7074-0.34830.7154-0.01240.73170.0167-0.23260.7478-0.11220.4964116.56841.7582353.2603
165.9222.63144.76484.25861.10174.16720.6286-2.37421.6041.6647-0.32653.2037-1.6256-3.5873-0.06860.68090.05040.37341.254-0.03820.929180.884347.6724327.2356
176.48040.62550.3592.9853-0.04175.7386-0.0525-0.2783-0.03130.21160.28090.11450.25280.0109-0.21450.31040.0029-0.01440.2136-0.0170.319993.347648.5844314.971
189.18685.7047-5.83213.5426-3.6233.71131.4569-0.4909-0.29970.7163-0.68590.5042-0.4701-1.8739-0.66741.1260.0511-0.17561.596-0.02270.7863119.646769.1006319.2728
192.5533-0.0331-0.0011.7629-0.74031.27750.034-0.15870.11040.053-0.0534-0.2478-0.03410.05410.03310.2758-0.0144-0.01860.2411-0.03950.2962105.5559.5732307.6187
204.46963.2655-2.4597.2342-5.43337.38460.01880.38160.3569-0.07230.64630.5835-0.0253-0.4809-0.80290.1752-0.01090.05470.3669-0.00070.405288.470264.0738302.5919
214.5566-1.85540.80845.4508-1.79835.0125-0.1629-0.0547-0.3086-0.35780.0016-0.22190.22920.44970.13320.3486-0.0670.05290.4247-0.04560.2822126.262328.5161252.8074
226.0398-1.2968-7.48477.25520.0459.6251-0.51445.9438-1.2328-4.2026-0.11051.40741.8743-2.31550.61751.8117-0.1814-0.23922.0172-0.13470.829994.951738.6557242.0545
232.7124-0.6454-1.0293.1578-0.65551.66190.01620.49250.0884-0.25050.10640.0305-0.238-0.1133-0.1410.4628-0.0903-0.03520.46770.0030.2476113.904642.4735246.9408
245.2845-0.6527-2.0519.6866.47844.91490.51122.0083-0.0643-1.0209-1.03490.61270.0858-1.99310.74830.8009-0.18570.00521.25470.00080.482111.425542.6485232.0624
253.4384-1.58161.62724.7606-3.72287.7577-0.02720.82610.2016-1.1993-0.033-0.7930.13710.6767-0.00710.8068-0.15520.16440.8426-0.10290.4223127.594835.6603231.9183
266.8187-2.21590.36463.8142-0.34654.01180.07740.38640.2449-0.1876-0.00690.1784-0.3192-0.1398-0.13470.3142-0.05820.02510.17120.01160.318396.031529.3903264.5424
276.2862-6.825-5.95887.43236.48015.65360.8148-0.0260.0655-1.2972-0.6863-0.0296-0.3166-0.9664-0.25730.88390.0857-0.02191.1382-0.140.6151121.562111.4672267.172
288.4374-2.4685-1.07493.7123-1.85473.86740.13420.24-0.0522-0.0119-0.0585-0.2442-0.08940.3431-0.07150.2272-0.0727-0.0180.2771-0.06360.246114.030624.4115268.9871
294.57290.2030.921.9897-0.2441.5664-0.06680.0228-0.0519-0.00520.04340.00350.0140.05350.01760.2779-0.01170.01030.1768-0.01020.2239103.02514.4262277.0487
305.58982.8715-5.21824.1694-1.39215.48920.6693-0.0592-1.084-0.0795-1.05870.68910.32080.45740.31150.57010.1066-0.03440.48680.05590.413695.0194-1.3792285.494
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1(CHAIN A AND RESID 4:66)
2X-RAY DIFFRACTION2(CHAIN A AND RESID 67:74)
3X-RAY DIFFRACTION3(CHAIN A AND RESID 75:179)
4X-RAY DIFFRACTION4(CHAIN A AND RESID 180:193)
5X-RAY DIFFRACTION5(CHAIN A AND RESID 194:262)
6X-RAY DIFFRACTION6(CHAIN B AND RESID 3:14)
7X-RAY DIFFRACTION7(CHAIN B AND RESID 15:67)
8X-RAY DIFFRACTION8(CHAIN B AND RESID 68:74)
9X-RAY DIFFRACTION9(CHAIN B AND RESID 75:210)
10X-RAY DIFFRACTION10(CHAIN B AND RESID 211:262)
11X-RAY DIFFRACTION11(CHAIN C AND RESID 3:50)
12X-RAY DIFFRACTION12(CHAIN C AND RESID 51:67)
13X-RAY DIFFRACTION13(CHAIN C AND RESID 68:88)
14X-RAY DIFFRACTION14(CHAIN C AND RESID 89:206)
15X-RAY DIFFRACTION15(CHAIN C AND RESID 207:263)
16X-RAY DIFFRACTION16(CHAIN D AND RESID 3:6)
17X-RAY DIFFRACTION17(CHAIN D AND RESID 7:69)
18X-RAY DIFFRACTION18(CHAIN D AND RESID 70:75)
19X-RAY DIFFRACTION19(CHAIN D AND RESID 76:227)
20X-RAY DIFFRACTION20(CHAIN D AND RESID 228:262)
21X-RAY DIFFRACTION21(CHAIN E AND RESID 3:68)
22X-RAY DIFFRACTION22(CHAIN E AND RESID 69:75)
23X-RAY DIFFRACTION23(CHAIN E AND RESID 76:199)
24X-RAY DIFFRACTION24(CHAIN E AND RESID 200:210)
25X-RAY DIFFRACTION25(CHAIN E AND RESID 211:263)
26X-RAY DIFFRACTION26(CHAIN F AND RESID 4:66)
27X-RAY DIFFRACTION27(CHAIN F AND RESID 67:72)
28X-RAY DIFFRACTION28(CHAIN F AND RESID 73:129)
29X-RAY DIFFRACTION29(CHAIN F AND RESID 130:256)
30X-RAY DIFFRACTION30(CHAIN F AND RESID 257:262)

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