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- PDB-5j32: Isopropylmalate dehydrogenase in complex with isopropylmalate -

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Basic information

Entry
Database: PDB / ID: 5j32
TitleIsopropylmalate dehydrogenase in complex with isopropylmalate
Components3-isopropylmalate dehydrogenase 2, chloroplastic
KeywordsOXIDOREDUCTASE / dehydrogenase / leucine biosynthesis / glucosinolate biosynthesis
Function / homology
Function and homology information


embryo sac development / 3-isopropylmalate dehydrogenase / 3-isopropylmalate dehydrogenase activity / pollen development / L-leucine biosynthetic process / chloroplast envelope / plastid / chloroplast stroma / NAD+ binding / amino acid biosynthetic process ...embryo sac development / 3-isopropylmalate dehydrogenase / 3-isopropylmalate dehydrogenase activity / pollen development / L-leucine biosynthetic process / chloroplast envelope / plastid / chloroplast stroma / NAD+ binding / amino acid biosynthetic process / chloroplast / magnesium ion binding / protein homodimerization activity / cytosol
Similarity search - Function
Isopropylmalate dehydrogenase / Isopropylmalate Dehydrogenase / Isopropylmalate Dehydrogenase / Isocitrate/isopropylmalate dehydrogenase, conserved site / Isocitrate and isopropylmalate dehydrogenases signature. / Isopropylmalate dehydrogenase-like domain / Isocitrate/isopropylmalate dehydrogenase / Isocitrate/isopropylmalate dehydrogenase / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
3-ISOPROPYLMALIC ACID / 3-isopropylmalate dehydrogenase 2, chloroplastic
Similarity search - Component
Biological speciesArabidopsis thaliana (thale cress)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.933 Å
AuthorsJez, J.M. / Lee, S.G.
Funding support United States, 1items
OrganizationGrant numberCountry
National Science Foundation (NSF, United States)MCB-0904215 United States
CitationJournal: J.Biol.Chem. / Year: 2016
Title: Structure and Mechanism of Isopropylmalate Dehydrogenase from Arabidopsis thaliana: INSIGHTS ON LEUCINE AND ALIPHATIC GLUCOSINOLATE BIOSYNTHESIS.
Authors: Lee, S.G. / Nwumeh, R. / Jez, J.M.
History
DepositionMar 30, 2016Deposition site: RCSB / Processing site: RCSB
Revision 1.0May 11, 2016Provider: repository / Type: Initial release
Revision 1.1May 18, 2016Group: Database references
Revision 1.2Jul 6, 2016Group: Database references
Revision 1.3Sep 27, 2017Group: Advisory / Author supporting evidence ...Advisory / Author supporting evidence / Database references / Derived calculations
Category: citation / pdbx_audit_support ...citation / pdbx_audit_support / pdbx_struct_oper_list / pdbx_unobs_or_zero_occ_atoms
Item: _citation.journal_id_CSD / _pdbx_audit_support.funding_organization / _pdbx_struct_oper_list.symmetry_operation
Revision 1.4Nov 27, 2019Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.5Sep 27, 2023Group: Advisory / Data collection ...Advisory / 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 / pdbx_struct_conn_angle / pdbx_unobs_or_zero_occ_atoms / struct_conn
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_struct_conn_angle.ptnr1_auth_asym_id / _pdbx_struct_conn_angle.ptnr1_auth_comp_id / _pdbx_struct_conn_angle.ptnr1_auth_seq_id / _pdbx_struct_conn_angle.ptnr1_label_asym_id / _pdbx_struct_conn_angle.ptnr1_label_atom_id / _pdbx_struct_conn_angle.ptnr1_label_comp_id / _pdbx_struct_conn_angle.ptnr1_label_seq_id / _pdbx_struct_conn_angle.ptnr3_auth_asym_id / _pdbx_struct_conn_angle.ptnr3_auth_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_seq_id / _pdbx_struct_conn_angle.ptnr3_label_asym_id / _pdbx_struct_conn_angle.ptnr3_label_atom_id / _pdbx_struct_conn_angle.ptnr3_label_comp_id / _pdbx_struct_conn_angle.ptnr3_label_seq_id / _pdbx_struct_conn_angle.value / _struct_conn.pdbx_dist_value / _struct_conn.ptnr1_auth_asym_id / _struct_conn.ptnr1_auth_comp_id / _struct_conn.ptnr1_auth_seq_id / _struct_conn.ptnr1_label_asym_id / _struct_conn.ptnr1_label_atom_id / _struct_conn.ptnr1_label_comp_id / _struct_conn.ptnr1_label_seq_id / _struct_conn.ptnr2_auth_asym_id / _struct_conn.ptnr2_auth_comp_id / _struct_conn.ptnr2_auth_seq_id / _struct_conn.ptnr2_label_asym_id / _struct_conn.ptnr2_label_atom_id / _struct_conn.ptnr2_label_comp_id / _struct_conn.ptnr2_label_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: 3-isopropylmalate dehydrogenase 2, chloroplastic
B: 3-isopropylmalate dehydrogenase 2, chloroplastic
C: 3-isopropylmalate dehydrogenase 2, chloroplastic
D: 3-isopropylmalate dehydrogenase 2, chloroplastic
hetero molecules


Theoretical massNumber of molelcules
Total (without water)173,61812
Polymers172,8164
Non-polymers8028
Water25,8341434
1
A: 3-isopropylmalate dehydrogenase 2, chloroplastic
B: 3-isopropylmalate dehydrogenase 2, chloroplastic
hetero molecules


Theoretical massNumber of molelcules
Total (without water)86,8096
Polymers86,4082
Non-polymers4014
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area6210 Å2
ΔGint-45 kcal/mol
Surface area27980 Å2
MethodPISA
2
C: 3-isopropylmalate dehydrogenase 2, chloroplastic
D: 3-isopropylmalate dehydrogenase 2, chloroplastic
hetero molecules


Theoretical massNumber of molelcules
Total (without water)86,8096
Polymers86,4082
Non-polymers4014
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area6040 Å2
ΔGint-45 kcal/mol
Surface area26810 Å2
MethodPISA
Unit cell
Length a, b, c (Å)96.059, 49.622, 158.969
Angle α, β, γ (deg.)90.00, 105.44, 90.00
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein
3-isopropylmalate dehydrogenase 2, chloroplastic / IMDH 2 / Beta-IPM dehydrogenase 2


Mass: 43204.086 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Arabidopsis thaliana (thale cress) / Gene: IMDH2, IMDH, At1g80560, T21F11.11 / Production host: Escherichia coli (E. coli)
References: UniProt: P93832, 3-isopropylmalate dehydrogenase
#2: Chemical
ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Formula: Mg
#3: Chemical
ChemComp-IPM / 3-ISOPROPYLMALIC ACID


Mass: 176.167 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C7H12O5
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1434 / 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: 2.1 Å3/Da / Density % sol: 41.48 %
Crystal growTemperature: 277 K / Method: vapor diffusion, hanging drop / pH: 8
Details: 1.0 M ammonium phosphate, 0.1 M imidazole, pH 8.0, and 5 mM isopropylmalate

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-ID / Wavelength: 0.979 Å
DetectorType: ADSC QUANTUM 315r / Detector: CCD / Date: Oct 12, 2012
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.979 Å / Relative weight: 1
ReflectionResolution: 1.93→32.5 Å / Num. obs: 104824 / % possible obs: 96.4 % / Observed criterion σ(F): 0 / Observed criterion σ(I): 0 / Redundancy: 3.15 % / Rsym value: 0.093 / Net I/σ(I): 11
Reflection shellResolution: 1.93→1.96 Å / Redundancy: 3.1 % / Rmerge(I) obs: 0.624 / Mean I/σ(I) obs: 2 / % possible all: 92.6

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Processing

Software
NameVersionClassification
PHENIX1.8.1_1168refinement
HKL-3000data reduction
HKL-3000data scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 3R8W
Resolution: 1.933→32.475 Å / SU ML: 0.18 / Cross valid method: FREE R-VALUE / σ(F): 1.35 / Phase error: 18.51 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.198 5246 5.01 %
Rwork0.1587 --
obs0.1607 104735 96.08 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 1.933→32.475 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms10948 0 52 1434 12434
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00711222
X-RAY DIFFRACTIONf_angle_d1.02115189
X-RAY DIFFRACTIONf_dihedral_angle_d13.3264201
X-RAY DIFFRACTIONf_chiral_restr0.0661746
X-RAY DIFFRACTIONf_plane_restr0.0051998
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.9328-1.95480.26971770.20452790X-RAY DIFFRACTION83
1.9548-1.97780.25261670.19253206X-RAY DIFFRACTION93
1.9778-2.00190.23191790.19233324X-RAY DIFFRACTION95
2.0019-2.02720.26331610.18853183X-RAY DIFFRACTION95
2.0272-2.05390.22941740.17743208X-RAY DIFFRACTION93
2.0539-2.0820.22822030.17653266X-RAY DIFFRACTION96
2.082-2.11180.23011750.16983270X-RAY DIFFRACTION97
2.1118-2.14330.21041630.17093249X-RAY DIFFRACTION94
2.1433-2.17680.21971730.16373314X-RAY DIFFRACTION97
2.1768-2.21240.21841810.163348X-RAY DIFFRACTION98
2.2124-2.25060.19921570.16473319X-RAY DIFFRACTION94
2.2506-2.29150.2091830.16173325X-RAY DIFFRACTION98
2.2915-2.33560.20222000.15433317X-RAY DIFFRACTION97
2.3356-2.38320.2061620.15973296X-RAY DIFFRACTION96
2.3832-2.4350.20671660.15563408X-RAY DIFFRACTION99
2.435-2.49170.19731760.15393335X-RAY DIFFRACTION96
2.4917-2.55390.22621790.15823387X-RAY DIFFRACTION100
2.5539-2.6230.21591700.16123367X-RAY DIFFRACTION96
2.623-2.70010.2021600.163422X-RAY DIFFRACTION100
2.7001-2.78720.18721890.15663347X-RAY DIFFRACTION97
2.7872-2.88680.20651660.15433429X-RAY DIFFRACTION99
2.8868-3.00230.1891640.15933393X-RAY DIFFRACTION98
3.0023-3.13880.19741950.14773339X-RAY DIFFRACTION97
3.1388-3.30410.1641630.13973421X-RAY DIFFRACTION98
3.3041-3.51090.19421740.13883385X-RAY DIFFRACTION98
3.5109-3.78160.16081840.1423374X-RAY DIFFRACTION97
3.7816-4.16150.18391750.13763369X-RAY DIFFRACTION96
4.1615-4.76210.15791910.13993317X-RAY DIFFRACTION96
4.7621-5.99350.17151760.17563373X-RAY DIFFRACTION95
5.9935-32.47920.21221630.18813408X-RAY DIFFRACTION93
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)
11.35960.07050.04430.8829-0.1381.70920.0466-0.20560.17150.10250.04340.0917-0.1754-0.0075-0.08690.1356-0.02230.03150.1349-0.01040.096718.83999.573435.9181
20.98470.23990.00570.6558-0.0960.78870.0119-0.0858-0.07150.038-0.0336-0.02320.07530.00220.01760.0818-0.00520.01840.0810.02880.084526.3769-3.679716.5068
31.1791.29180.30592.6256-0.1922.3037-0.13410.23770.4104-0.10150.01650.5299-0.2509-0.14260.00190.1282-0.01560.02830.11050.03730.17297.49248.265425.4838
42.18571.06050.68650.73930.27881.25820.0988-0.2021-0.43720.1559-0.0444-0.03080.16020.0711-0.00560.1667-0.03310.06980.14870.05090.153712.0691-6.796334.8097
51.4459-0.1287-0.05041.9207-0.13493.23360.0335-0.2630.020.1709-0.15010.3004-0.0776-0.35070.0480.1035-0.01530.05860.22410.00150.1734-2.7243-1.492129.2475
61.67180.10190.34121.54620.27330.8470.05220.1137-0.1077-0.08790.0446-0.1190.04350.1588-0.02760.09380.00540.07380.11340.02320.086535.9227-3.9421-16.813
70.64140.03750.03490.6699-0.12940.85440.0026-0.00130.0516-0.0054-0.00890.0302-0.0646-0.00590.00520.0665-0.00560.02380.06390.02030.079523.02797.84850.4631
81.5574-0.23740.1632.6755-0.60662.24960.0218-0.1075-0.33050.15510.0154-0.08620.18530.0659-0.02250.0961-0.00970.00580.09520.0310.135625.7143-13.0217-10.8299
92.26211.70190.19522.0322-0.31061.1647-0.01810.16120.3431-0.15870.10590.474-0.0968-0.0945-0.09730.0960.0198-0.00160.10530.01160.126218.4281-0.1801-21.5297
102.42350.2568-0.18073.2403-1.33363.36860.02150.2371-0.2497-0.28040.10520.06850.2464-0.184-0.11670.108-0.0221-0.03090.0932-0.01050.122113.6495-15.4787-19.2714
111.7177-0.61830.37161.76120.42610.98040.05960.2203-0.0772-0.1914-0.0390.01580.04570.10750.00410.15140.00770.05410.15950.00970.087111.16156.892663.9835
121.1352-0.13470.02930.9223-0.13180.90610.00110.08770.1127-0.003-0.0473-0.0714-0.17780.09580.03430.1481-0.02610.01920.09820.03580.09919.293421.808882.9807
133.14890.6147-0.2713.8482-0.48852.9519-0.0087-0.083-0.2428-0.2695-0.16960.3549-0.0252-0.15050.14090.1290.0469-0.00410.1235-0.01410.132-2.81310.223665.5609
142.66890.49310.18540.09580.03561.1890.09510.05490.5769-0.2928-0.1768-0.1564-0.33560.08570.02470.27620.0060.04440.16170.07270.19996.208225.109560
151.27060.3984-0.38872.6403-0.73482.71560.09060.41880.0437-0.3908-0.14360.4057-0.0381-0.25650.06250.23260.0599-0.11150.28260.00090.2007-10.506119.778958.1289
161.154-0.40220.4561.45090.05391.6568-0.08750.06210.12610.14730.0785-0.0942-0.04310.1014-0.01510.1446-0.0040.01230.11970.02280.0861.057622.1909116.3212
170.88670.0367-0.05980.47970.01330.7828-0.0431-0.0657-0.0612-0.02160.01880.02740.0083-0.06040.02420.1298-0.00160.01790.07940.02080.0797-1.712110.271595.1167
182.31880.00860.2970.83670.1461.2262-0.1496-0.01110.3838-0.1339-0.0206-0.2355-0.15140.10280.11050.1759-0.0190.0080.13030.06130.1407-5.144831.2629106.4195
191.2241-0.97850.13411.7031-1.33851.65560.1937-0.2049-0.3783-0.22110.08820.46010.3079-0.3139-0.25160.1356-0.0272-0.01230.17110.06420.1867-16.914418.251111.5848
202.60040.16580.31452.7593-1.01873.753-0.0419-0.13980.26620.30260.08230.1993-0.2411-0.3803-0.02750.1580.0330.04820.14880.03480.1815-19.09734.363106.1833
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1CHAIN A AND RESID 31:142
2X-RAY DIFFRACTION2CHAIN A AND RESID 143:316
3X-RAY DIFFRACTION3CHAIN A AND RESID 317:337
4X-RAY DIFFRACTION4CHAIN A AND RESID 338:367
5X-RAY DIFFRACTION5CHAIN A AND RESID 368:399
6X-RAY DIFFRACTION6CHAIN B AND RESID 36:142
7X-RAY DIFFRACTION7CHAIN B AND RESID 143:316
8X-RAY DIFFRACTION8CHAIN B AND RESID 317:337
9X-RAY DIFFRACTION9CHAIN B AND RESID 338:367
10X-RAY DIFFRACTION10CHAIN B AND RESID 368:400
11X-RAY DIFFRACTION11CHAIN C AND RESID 41:142
12X-RAY DIFFRACTION12CHAIN C AND RESID 143:316
13X-RAY DIFFRACTION13CHAIN C AND RESID 317:337
14X-RAY DIFFRACTION14CHAIN C AND RESID 338:367
15X-RAY DIFFRACTION15CHAIN C AND RESID 368:396
16X-RAY DIFFRACTION16CHAIN D AND RESID 38:142
17X-RAY DIFFRACTION17CHAIN D AND RESID 143:316
18X-RAY DIFFRACTION18CHAIN D AND RESID 317:337
19X-RAY DIFFRACTION19CHAIN D AND RESID 338:367
20X-RAY DIFFRACTION20CHAIN D AND RESID 368:397

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