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- PDB-5uqq: Crystal structure of 2-methylcitrate synthase from Aspergillus fu... -

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Basic information

Entry
Database: PDB / ID: 5uqq
TitleCrystal structure of 2-methylcitrate synthase from Aspergillus fumigatus
Components2-methylcitrate synthase, mitochondrial
KeywordsTRANSFERASE / mcsA / 2-methylcitrate synthase / citrate synthase
Function / homology
Function and homology information


2-methylcitrate synthase / 2-methylcitrate synthase activity / citrate synthase (unknown stereospecificity) / propionate catabolic process / citrate (Si)-synthase activity / tricarboxylic acid cycle / carbohydrate metabolic process / mitochondrial matrix
Similarity search - Function
Citrate Synthase; domain 1 / Citrate Synthase, domain 1 / Cytochrome p450-Terp; domain 2 / Cytochrome P450-Terp, domain 2 / Citrate synthase active site / Citrate synthase signature. / Citrate synthase-like, large alpha subdomain / Citrate synthase / Citrate synthase-like, small alpha subdomain / Citrate synthase superfamily ...Citrate Synthase; domain 1 / Citrate Synthase, domain 1 / Cytochrome p450-Terp; domain 2 / Cytochrome P450-Terp, domain 2 / Citrate synthase active site / Citrate synthase signature. / Citrate synthase-like, large alpha subdomain / Citrate synthase / Citrate synthase-like, small alpha subdomain / Citrate synthase superfamily / Citrate synthase, C-terminal domain / Orthogonal Bundle / Mainly Alpha
Similarity search - Domain/homology
S,R MESO-TARTARIC ACID / 2-methylcitrate synthase, mitochondrial
Similarity search - Component
Biological speciesNeosartorya fumigata (mold)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.3 Å
AuthorsSchlachter, C. / Chruszcz, M.
CitationJournal: Biol.Chem. / Year: 2019
Title: Comparative studies of Aspergillus fumigatus 2-methylcitrate synthase and human citrate synthase.
Authors: Schlachter, C.R. / Klapper, V. / Radford, T. / Chruszcz, M.
History
DepositionFeb 8, 2017Deposition site: RCSB / Processing site: RCSB
Revision 1.0Feb 14, 2018Provider: repository / Type: Initial release
Revision 1.1Jun 17, 2020Group: 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
Revision 1.2Mar 6, 2024Group: Advisory / Data collection ...Advisory / Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_unobs_or_zero_occ_atoms / struct_ncs_dom_lim
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.beg_label_asym_id / _struct_ncs_dom_lim.beg_label_comp_id / _struct_ncs_dom_lim.beg_label_seq_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_ncs_dom_lim.end_label_asym_id / _struct_ncs_dom_lim.end_label_comp_id / _struct_ncs_dom_lim.end_label_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
B: 2-methylcitrate synthase, mitochondrial
C: 2-methylcitrate synthase, mitochondrial
A: 2-methylcitrate synthase, mitochondrial
D: 2-methylcitrate synthase, mitochondrial
E: 2-methylcitrate synthase, mitochondrial
F: 2-methylcitrate synthase, mitochondrial
hetero molecules


Theoretical massNumber of molelcules
Total (without water)306,3257
Polymers306,1746
Non-polymers1501
Water26,1041449
1
B: 2-methylcitrate synthase, mitochondrial
C: 2-methylcitrate synthase, mitochondrial
hetero molecules


Theoretical massNumber of molelcules
Total (without water)102,2083
Polymers102,0582
Non-polymers1501
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area8920 Å2
ΔGint-67 kcal/mol
Surface area31900 Å2
MethodPISA
2
A: 2-methylcitrate synthase, mitochondrial
D: 2-methylcitrate synthase, mitochondrial


Theoretical massNumber of molelcules
Total (without water)102,0582
Polymers102,0582
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area8450 Å2
ΔGint-67 kcal/mol
Surface area31180 Å2
MethodPISA
3
E: 2-methylcitrate synthase, mitochondrial
F: 2-methylcitrate synthase, mitochondrial


Theoretical massNumber of molelcules
Total (without water)102,0582
Polymers102,0582
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area8580 Å2
ΔGint-62 kcal/mol
Surface area31400 Å2
MethodPISA
Unit cell
Length a, b, c (Å)82.393, 130.012, 261.535
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11B
21C
12B
22A
13B
23D
14B
24E
15B
25F
16C
26A
17C
27D
18C
28E
19C
29F
110A
210D
111A
211E
112A
212F
113D
213E
114D
214F
115E
215F

NCS domain segments:

Component-ID: _ / Refine code: _

Dom-IDEns-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11THRTHRLYSLYSBA30 - 46427 - 461
21THRTHRLYSLYSCB30 - 46427 - 461
12ALAALALYSLYSBA31 - 46428 - 461
22ALAALALYSLYSAC31 - 46428 - 461
13ALAALALEULEUBA31 - 46328 - 460
23ALAALALEULEUDD31 - 46328 - 460
14ALAALALEULEUBA31 - 46328 - 460
24ALAALALEULEUEE31 - 46328 - 460
15ALAALALYSLYSBA31 - 46428 - 461
25ALAALALYSLYSFF31 - 46428 - 461
16ALAALALYSLYSCB31 - 46428 - 461
26ALAALALYSLYSAC31 - 46428 - 461
17ALAALALEULEUCB31 - 46328 - 460
27ALAALALEULEUDD31 - 46328 - 460
18ALAALALEULEUCB31 - 46328 - 460
28ALAALALEULEUEE31 - 46328 - 460
19ALAALALYSLYSCB31 - 46428 - 461
29ALAALALYSLYSFF31 - 46428 - 461
110ALAALALEULEUAC31 - 46328 - 460
210ALAALALEULEUDD31 - 46328 - 460
111ALAALALEULEUAC31 - 46328 - 460
211ALAALALEULEUEE31 - 46328 - 460
112ALAALALYSLYSAC31 - 46528 - 462
212ALAALALYSLYSFF31 - 46528 - 462
113ALAALALYSLYSDD31 - 46428 - 461
213ALAALALYSLYSEE31 - 46428 - 461
114ALAALALEULEUDD31 - 46328 - 460
214ALAALALEULEUFF31 - 46328 - 460
115ALAALALEULEUEE31 - 46328 - 460
215ALAALALEULEUFF31 - 46328 - 460

NCS ensembles :
ID
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15

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Components

#1: Protein
2-methylcitrate synthase, mitochondrial / Methylcitrate synthase / (2S / 3S)-2-methylcitrate synthase / Citrate synthase 1


Mass: 51029.078 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Neosartorya fumigata (mold) / Gene: mcsA, cit1 / Plasmid: pJExpress411 / Cell line (production host): BL-21(DE3) / Production host: Escherichia coli (E. coli)
References: UniProt: Q50I20, 2-methylcitrate synthase, citrate synthase (unknown stereospecificity)
#2: Chemical ChemComp-SRT / S,R MESO-TARTARIC ACID


Mass: 150.087 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C4H6O6
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1449 / 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: 2.29 Å3/Da / Density % sol: 46.23 %
Crystal growTemperature: 298 K / Method: vapor diffusion, sitting drop / Details: 0.2 M NaTartrate, 20% PEG3350 pH 7.85

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 22-BM / Wavelength: 1 Å
DetectorType: MARMOSAIC 225 mm CCD / Detector: CCD / Date: Oct 16, 2013
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 2.3→50 Å / Num. obs: 118739 / % possible obs: 99.67 % / Observed criterion σ(I): -3 / Redundancy: 4.9 % / Rmerge(I) obs: 0.081 / Rpim(I) all: 0.052 / Rrim(I) all: 0.117 / Rsym value: 0.081 / Net I/σ(I): 18.4
Reflection shellResolution: 2.3→2.33 Å / Redundancy: 4.8 % / Rmerge(I) obs: 0.528 / Mean I/σ(I) obs: 2.35 / Num. unique obs: 4770 / CC1/2: 0.809 / Rpim(I) all: 0.354 / Rrim(I) all: 0.777 / Rsym value: 0.528 / % possible all: 99.7

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Processing

Software
NameVersionClassification
REFMAC5.8.0158refinement
HKL-2000data scaling
MOLREPphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.3→31.41 Å / Cor.coef. Fo:Fc: 0.956 / Cor.coef. Fo:Fc free: 0.941 / Cross valid method: THROUGHOUT / ESU R: 0.36 / ESU R Free: 0.215 / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.21434 6278 5 %RANDOM
Rwork0.18524 ---
obs0.1867 118739 99.67 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å
Displacement parametersBiso mean: 42.189 Å2
Baniso -1Baniso -2Baniso -3
1--0.45 Å2-0 Å20 Å2
2--0.34 Å2-0 Å2
3---0.11 Å2
Refinement stepCycle: 1 / Resolution: 2.3→31.41 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms20274 0 10 1449 21733
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0120.01920832
X-RAY DIFFRACTIONr_bond_other_d00.0219572
X-RAY DIFFRACTIONr_angle_refined_deg1.5091.9728286
X-RAY DIFFRACTIONr_angle_other_deg3.641345441
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.2752616
X-RAY DIFFRACTIONr_dihedral_angle_2_deg38.05424.118896
X-RAY DIFFRACTIONr_dihedral_angle_3_deg14.511153497
X-RAY DIFFRACTIONr_dihedral_angle_4_deg17.67315113
X-RAY DIFFRACTIONr_chiral_restr0.0850.23137
X-RAY DIFFRACTIONr_gen_planes_refined0.0080.02123179
X-RAY DIFFRACTIONr_gen_planes_other0.0110.024138
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_it1.1022.19710465
X-RAY DIFFRACTIONr_mcbond_other1.1022.19710464
X-RAY DIFFRACTIONr_mcangle_it1.9253.2913079
X-RAY DIFFRACTIONr_mcangle_other1.9253.2913080
X-RAY DIFFRACTIONr_scbond_it1.2732.3710367
X-RAY DIFFRACTIONr_scbond_other1.2732.36910364
X-RAY DIFFRACTIONr_scangle_it
X-RAY DIFFRACTIONr_scangle_other2.2253.47315205
X-RAY DIFFRACTIONr_long_range_B_refined5.28141.16488080
X-RAY DIFFRACTIONr_long_range_B_other5.28141.16288079
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION / Type: interatomic distance / Weight position: 0.05

Ens-IDDom-IDAuth asym-IDNumberRms dev position (Å)
11B278560.07
12C278560.07
21B279920.07
22A279920.07
31B276640.08
32D276640.08
41B278480.07
42E278480.07
51B275640.08
52F275640.08
61C278460.07
62A278460.07
71C277460.07
72D277460.07
81C278620.07
82E278620.07
91C277100.07
92F277100.07
101A276280.07
102D276280.07
111A279240.06
112E279240.06
121A278240.07
122F278240.07
131D277480.08
132E277480.08
141D277720.07
142F277720.07
151E275720.07
152F275720.07
LS refinement shellResolution: 2.3→2.359 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.281 455 -
Rwork0.258 8584 -
obs--98.84 %
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)
10.65750.11180.05550.2419-0.05130.74970.01770.0479-0.0807-0.00830.0360.01390.0916-0.1725-0.05380.1302-0.0149-0.00430.18090.01380.1657-6.545-49.75620.668
22.8169-0.3502-0.19370.64841.02855.6680.0211-0.48130.5517-0.30620.0881-0.2725-0.82560.1173-0.10920.30450.05790.09720.1847-0.050.2325-17.561-19.61923.066
30.6007-0.10050.10060.03670.09520.66240.030.04280.0039-0.0281-0.0474-0.0046-0.1242-0.19510.01730.15410.03890.01850.22540.01980.1411-11.721-34.63319.017
42.5675-0.8002-1.20680.98850.46571.8101-0.1365-0.1905-0.49120.12330.10540.13260.22980.11740.03110.16920.0026-0.0530.09960.05730.1511-11.209-35.76271.71
50.5638-0.22340.30840.61470.09050.67130.00710.0969-0.0448-0.04260.0609-0.0290.03830.1131-0.06810.1526-0.0083-0.02170.1827-0.02310.1355-0.937-25.68658.135
60.356-0.0490.14890.55860.37360.3941-0.02580.064400.02680.1357-0.07980.01430.1166-0.10990.1515-0.0043-0.01990.2066-0.03480.15285.933-15.17870.027
70.72450.12480.02320.7946-0.12680.6586-0.0411-0.07150.14190.05480.00180.1131-0.13430.02460.03930.1802-0.0004-0.03840.1025-0.02570.1714-25.394-4.79165.248
81.21230.50320.75361.33121.17872.13830.02590.07330.03470.1479-0.09970.01930.2232-0.09990.07370.13140.0119-0.01170.1415-0.0290.1614-44.923-23.85657.664
90.82160.67550.16390.71420.12370.1836-0.054-0.01770.07490.0959-0.06940.0715-0.04020.06680.12350.20580.0146-0.00410.172800.1291-18.663-16.02670.062
100.69390.0107-0.05770.25680.00621.00690.0133-0.03130.07680.01750.014-0.0233-0.09350.072-0.02720.14-0.00280.00360.14050.0010.160321.013-35.38524.576
111.7942-0.07070.42812.398-0.35621.4888-0.01770.1384-0.1693-0.2743-0.0271-0.0970.12910.19610.04480.12120.03740.03220.1548-0.03610.156735.359-56.30112.315
120.55720.0745-0.24940.59390.3040.7868-0.00370.07030.0826-0.01910.0183-0.0379-0.06720.0025-0.01460.1230.00410.01090.16660.02340.148611.494-38.47116.355
130.56-0.29250.18310.4869-0.33430.74660.1202-0.0393-0.2839-0.42010.04780.41710.1638-0.0417-0.1680.5045-0.0688-0.39780.02230.05050.3939-32.51616.16925.525
142.3326-0.3443-0.24333.7261-1.23061.1220.5382-0.1848-0.2406-0.4543-0.06680.4033-0.0031-0.017-0.47140.5381-0.0004-0.21010.09690.01990.3413-46.03540.87423.247
150.4811-0.41790.33491.2527-0.57350.64720.15410.0031-0.2296-0.5540.00860.46320.00550.0615-0.16270.6324-0.0696-0.35260.05860.01670.253-23.69722.16118.336
160.73620.11530.18090.69220.32820.48030.04190.0653-0.0144-0.36460.05110.0768-0.07240.0989-0.09290.4652-0.0496-0.11120.0654-0.0060.0928-4.75430.45224.952
171.5252-0.76981.89743.7167-3.40098.88660.00020.3235-0.5086-0.17040.1538-0.43830.00710.2171-0.1540.1603-0.03490.01080.1959-0.2680.41795.572.38513.894
181.46540.41570.12991.52440.55770.2489-0.16630.4166-0.3091-0.58080.288-0.2377-0.25230.1529-0.12170.5354-0.1051-0.01280.2271-0.17210.1719-0.57117.13917.637
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A31 - 309
2X-RAY DIFFRACTION2A310 - 355
3X-RAY DIFFRACTION3A356 - 465
4X-RAY DIFFRACTION4B30 - 75
5X-RAY DIFFRACTION5B76 - 217
6X-RAY DIFFRACTION6B218 - 465
7X-RAY DIFFRACTION7C29 - 310
8X-RAY DIFFRACTION8C311 - 422
9X-RAY DIFFRACTION9C423 - 465
10X-RAY DIFFRACTION10D31 - 311
11X-RAY DIFFRACTION11D312 - 422
12X-RAY DIFFRACTION12D423 - 464
13X-RAY DIFFRACTION13E31 - 310
14X-RAY DIFFRACTION14E311 - 422
15X-RAY DIFFRACTION15E423 - 464
16X-RAY DIFFRACTION16F31 - 311
17X-RAY DIFFRACTION17F312 - 357
18X-RAY DIFFRACTION18F358 - 465

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