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- PDB-9q6g: PYCR2 complexed with NAD+ and (2S)-2-hydroxy-3,3-dimethylbutanoic acid -

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Basic information

Entry
Database: PDB / ID: 9q6g
TitlePYCR2 complexed with NAD+ and (2S)-2-hydroxy-3,3-dimethylbutanoic acid
ComponentsPyrroline-5-carboxylate reductase 2
KeywordsOXIDOREDUCTASE / AMINO-ACID BIOSYNTHESIS / PROLINE BIOSYNTHESIS / BIOSYNTHESIS
Function / homology
Function and homology information


pyrroline-5-carboxylate reductase / pyrroline-5-carboxylate reductase activity / Glutamate and glutamine metabolism / L-glutamine catabolic process / L-proline biosynthetic process / L-glutamate catabolic process / cellular response to oxidative stress / mitochondrial matrix / mitochondrion / cytosol / cytoplasm
Similarity search - Function
: / Delta 1-pyrroline-5-carboxylate reductase signature. / Pyrroline-5-carboxylate reductase / Pyrroline-5-carboxylate reductase, dimerisation domain / Pyrroline-5-carboxylate reductase dimerisation / Pyrroline-5-carboxylate reductase, catalytic, N-terminal / NADP oxidoreductase coenzyme F420-dependent / 6-phosphogluconate dehydrogenase-like, C-terminal domain superfamily / NAD(P)-binding domain superfamily
Similarity search - Domain/homology
: / NICOTINAMIDE-ADENINE-DINUCLEOTIDE / Pyrroline-5-carboxylate reductase 2
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.65 Å
AuthorsTanner, J.J. / Meeks, K.R.
Funding support United States, 1items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)1R01GM132640 United States
CitationJournal: Acs Chem.Biol. / Year: 2026
Title: Targeting a Unique Cysteine Residue to Achieve Isoform-Selective Inhibition of the Proline Biosynthetic Enzyme Pyrroline-5-Carboxylate Reductase 2.
Authors: Rossman, T.C. / Meeks, K.R. / Purohit, G. / Naldrett, M.J. / Tanner, J.J. / Becker, D.F.
History
DepositionAug 22, 2025Deposition site: RCSB / Processing site: RCSB
Revision 1.0May 20, 2026Provider: repository / Type: Initial release
Revision 1.1May 27, 2026Group: Database references / Category: citation / citation_author
Item: _citation.journal_volume / _citation.page_first ..._citation.journal_volume / _citation.page_first / _citation.page_last / _citation_author.identifier_ORCID
Revision 1.2Sep 30, 2026Group: Structure summary / Category: struct_keywords / Item: _struct_keywords.text

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Pyrroline-5-carboxylate reductase 2
B: Pyrroline-5-carboxylate reductase 2
C: Pyrroline-5-carboxylate reductase 2
D: Pyrroline-5-carboxylate reductase 2
E: Pyrroline-5-carboxylate reductase 2
hetero molecules


Theoretical massNumber of molelcules
Total (without water)171,88314
Polymers168,1095
Non-polymers3,7749
Water1629
1
A: Pyrroline-5-carboxylate reductase 2
B: Pyrroline-5-carboxylate reductase 2
C: Pyrroline-5-carboxylate reductase 2
D: Pyrroline-5-carboxylate reductase 2
E: Pyrroline-5-carboxylate reductase 2
hetero molecules

A: Pyrroline-5-carboxylate reductase 2
B: Pyrroline-5-carboxylate reductase 2
C: Pyrroline-5-carboxylate reductase 2
D: Pyrroline-5-carboxylate reductase 2
E: Pyrroline-5-carboxylate reductase 2
hetero molecules


Theoretical massNumber of molelcules
Total (without water)343,76528
Polymers336,21810
Non-polymers7,54718
Water1086
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
point symmetry operation2_555-x,y,-z1
Buried area67440 Å2
ΔGint-668 kcal/mol
Surface area92610 Å2
MethodPISA
Unit cell
Length a, b, c (Å)104.336, 109.936, 154.004
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number18
Space group name H-MP21221

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Components

#1: Protein
Pyrroline-5-carboxylate reductase 2 / P5C reductase 2 / P5CR 2


Mass: 33621.801 Da / Num. of mol.: 5 / Mutation: K276Q
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: PYCR2 / Production host: Escherichia coli (E. coli)
References: UniProt: Q96C36, pyrroline-5-carboxylate reductase
#2: Chemical ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: SO4
#3: Chemical
ChemComp-NAD / NICOTINAMIDE-ADENINE-DINUCLEOTIDE


Mass: 663.425 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: C21H27N7O14P2 / Feature type: SUBJECT OF INVESTIGATION / Comment: NAD*YM
#4: Chemical ChemComp-G8I / (2S)-2-hydroxy-3,3-dimethylbutanoic acid


Mass: 132.158 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C6H12O3 / Feature type: SUBJECT OF INVESTIGATION
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 9 / Source method: isolated from a natural source / Formula: H2O
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: 2.63 Å3/Da / Density % sol: 53.18 %
Crystal growTemperature: 293 K / Method: vapor diffusion, sitting drop
Details: Resevoir contained 400 mM Li2SO4, 22% (w/v) PEG 3350, and 0.1 mM HEPES at pH 7.5. Enzyme solution contained 8 mM NAD+ and 10 mM 2S-Hydroxy-3,3-dimethylbutyric acid. Crystal was soaked in ...Details: Resevoir contained 400 mM Li2SO4, 22% (w/v) PEG 3350, and 0.1 mM HEPES at pH 7.5. Enzyme solution contained 8 mM NAD+ and 10 mM 2S-Hydroxy-3,3-dimethylbutyric acid. Crystal was soaked in cryobuffer containing 20% PEG 200.

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: ALS / Beamline: 5.0.3 / Wavelength: 0.976 Å
DetectorType: DECTRIS PILATUS3 2M / Detector: PIXEL / Date: Oct 26, 2024
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.976 Å / Relative weight: 1
ReflectionResolution: 2.65→49.41 Å / Num. obs: 99172 / % possible obs: 100 % / Redundancy: 6.8 % / CC1/2: 0.988 / Rmerge(I) obs: 0.304 / Rpim(I) all: 0.126 / Rrim(I) all: 0.329 / Χ2: 1.03 / Net I/σ(I): 7.6
Reflection shellResolution: 2.65→2.73 Å / Redundancy: 7.1 % / Rmerge(I) obs: 1.851 / Num. unique obs: 4479 / CC1/2: 0.348 / Rpim(I) all: 0.746 / Rrim(I) all: 1.998 / Χ2: 1.01

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Processing

Software
NameVersionClassification
PHENIX(1.21.2_5419: ???)refinement
Aimlessdata scaling
PDB_EXTRACTdata extraction
XDSdata reduction
PHASER1.21.2-5419phasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.65→48.63 Å / SU ML: 0.48 / Cross valid method: FREE R-VALUE / σ(F): 0.21 / Phase error: 34.1 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2843 1777 1.79 %
Rwork0.2683 --
obs0.2686 99172 99.74 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.1 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.65→48.63 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms8847 0 223 9 9079
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0029204
X-RAY DIFFRACTIONf_angle_d0.59512583
X-RAY DIFFRACTIONf_dihedral_angle_d13.243189
X-RAY DIFFRACTIONf_chiral_restr0.0421509
X-RAY DIFFRACTIONf_plane_restr0.0061661
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.65-2.720.42611260.3777500X-RAY DIFFRACTION100
2.72-2.80.37521480.37187515X-RAY DIFFRACTION100
2.8-2.890.36151150.35787503X-RAY DIFFRACTION100
2.89-30.4950.35457553X-RAY DIFFRACTION100
3-3.120.36721460.33287458X-RAY DIFFRACTION100
3.12-3.260.31731340.30847507X-RAY DIFFRACTION100
3.26-3.430.32861010.29047532X-RAY DIFFRACTION99
3.43-3.640.26951670.28397416X-RAY DIFFRACTION99
3.64-3.920.2921170.2487529X-RAY DIFFRACTION100
3.93-4.320.2571560.22477491X-RAY DIFFRACTION100
4.32-4.940.2581660.21847429X-RAY DIFFRACTION100
4.94-6.230.24481630.25237476X-RAY DIFFRACTION100
6.23-48.630.21571430.20227486X-RAY DIFFRACTION100
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL11 (°2)L12 (°2)L13 (°2)L22 (°2)L23 (°2)L33 (°2)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T11 (Å2)T12 (Å2)T13 (Å2)T22 (Å2)T23 (Å2)T33 (Å2)Origin x (Å)Origin y (Å)Origin z (Å)
12.8274-0.65790.62312.1089-0.98772.63220.15491.0771-0.5348-0.56410.02330.07781.0417-0.4197-0.16060.85330.0695-0.16460.7897-0.15360.6089-5.822127.8453-46.2356
20.68150.2936-0.34191.0964-0.31480.20070.03121.0808-0.7877-0.58990.02660.07831.0999-0.328-0.08651.2130.0362-0.30321.0181-0.41080.8899-6.761122.4348-50.0027
30.60860.25070.74311.5312-0.49221.3540.00451.3954-0.5777-0.91230.0349-0.15581.1319-0.1964-0.00181.3550.1376-0.15771.1973-0.40640.7964-0.533721.6544-54.7331
42.72540.0922-1.23712.2715-0.671.27710.1661.1882-0.5531-0.725-0.1725-0.34111.02690.23750.0270.74570.2718-0.0880.7493-0.19650.5638.64928.3498-47.6961
51.22120.0687-0.7750.0058-0.04240.49270.13390.9481-0.2556-0.5762-0.189-0.63250.77250.58710.05670.69420.36190.03450.9848-0.1330.478516.904233.5625-48.5862
62.29590.2306-0.61250.9702-0.45671.63540.29410.35880.0467-0.1197-0.0745-0.00580.5849-0.2089-0.19030.3146-0.0707-0.0650.26950.05270.2674-8.38836.7904-27.4932
72.16790.91610.5252.7868-0.92743.02530.29090.0384-0.3592-0.2362-0.2687-0.38780.76750.655-0.06090.45030.0883-0.1050.29610.03430.35066.621830.4907-22.2943
82.1106-0.38870.44140.7539-0.57230.89850.00690.00240.2730.09280.05830.16680.0567-0.3274-0.11820.2611-0.2057-0.06470.82440.18110.4576-38.998452.4308-23.9527
92.26060.4547-0.5391.62420.1450.7551-0.0124-0.3234-0.0837-0.00780.17310.33470.0215-0.5951-0.15970.4108-0.2616-0.01681.00330.22240.5904-46.323143.3839-18.7158
101.8414-0.05730.24990.5536-0.21460.26940.05810.0429-0.12260.00550.10050.27990.3089-0.4165-0.00050.2725-0.2626-0.02190.60750.11240.3982-29.599737.1945-20.0067
112.4682-0.23090.13771.0702-0.50661.00920.21750.0845-0.2562-0.2419-0.06140.21470.6819-0.3209-0.08880.6278-0.1698-0.0840.27840.04930.2799-11.703729.5636-19.8395
121.4066-0.291-0.35023.3799-0.90630.90550.25640.0326-0.2842-0.0281-0.0914-0.14940.79460.1485-0.18682.18080.1165-0.33040.56320.02650.68687.7597-16.965-19.177
130.1335-0.05840.18011.5411-0.27190.26770.24580.145-0.157-0.3028-0.11930.00270.5790.057-0.13862.26120.2622-0.32520.70740.12070.875813.6453-20.9332-17.4916
140.6795-0.2736-0.01541.02190.44650.75790.21370.2314-0.2471-0.2425-0.0575-0.22770.52110.217-0.0822.04580.5359-0.15390.7104-0.0830.586220.8991-8.3639-28.1938
150.4-0.0950.13820.2891-0.13360.09090.25010.2209-0.2439-0.3382-0.11170.04340.77670.0105-0.06081.8063-0.0941-0.27630.2263-0.09370.3869-4.70112.9391-19.0741
160.25970.00460.57111.4377-1.04882.03560.24640.1531-0.1699-0.4095-0.1907-0.18980.90330.3974-0.0261.31890.1898-0.21120.36980.05840.38279.150611.8359-16.1494
170.33730.13-0.69540.0634-0.27341.47660.19030.0516-0.1364-0.0971-0.18040.22580.4331-0.5634-0.00051.7655-0.2726-0.40631.0593-0.04880.9369-28.9626-0.1008-22.0663
180.49980.14840.02251.1166-1.57842.38060.19960.2014-0.1275-0.2938-0.22920.49170.3142-0.95840.02691.7891-0.2552-0.48681.1946-0.0840.91-33.6150.808-27.1604
190.3685-0.3355-0.50950.93680.10880.90230.16560.3543-0.4076-0.177-0.04360.51790.5292-0.8831-0.06781.5474-0.4797-0.46381.1645-0.01041.0235-40.94822.1841-12.1378
200.0046-0.0111-0.01360.0864-0.00010.0580.02470.5676-0.7467-0.28770.06380.6750.8828-1.047-0.05431.8426-0.6178-0.40471.4906-0.05031.2958-43.8886-2.1876-1.8126
210.2664-0.09250.17911.52780.52590.6710.19660.0891-0.34920.0183-0.24630.2590.4033-0.60440.07721.5233-0.4621-0.40670.63350.03730.59-25.6358-4.6536-8.1566
220.1133-0.10090.08890.30230.18990.39680.06470.0071-0.1066-0.1039-0.07580.06530.472-0.036-0.17821.50360.03-0.30280.2284-0.09330.3447-0.7255.1047-14.2767
230.75920.5355-0.40711.5841-0.69181.26220.1237-0.00470.03420.00980.08410.3480.3267-0.3018-0.29781.2002-0.3717-0.20260.38340.12430.4026-19.411512.7065-1.6348
242.9642-0.5596-1.17830.4789-0.56272.15090.18170.78080.6797-0.2688-0.1116-0.072-0.34920.0111-0.1590.51950.08550.05140.68830.19580.72058.244767.4374-22.4862
252.7224-0.259-0.74560.0470.23411.39920.11320.89250.5815-0.3483-0.0825-0.1878-0.38910.1051-0.16650.39980.07340.17610.87310.14290.632518.244162.9517-26.815
261.5520.1593-0.38690.5564-0.00170.91660.08890.09530.3449-0.008-0.1042-0.1669-0.19270.2924-0.0733-0.1113-0.0227-0.01080.31690.05650.49759.785856.7634-4.9044
271.3486-0.23030.97422.10260.44080.88240.08880.2931-0.0788-0.2367-0.0585-0.4660.10630.45080.00150.16220.1442-0.09820.4259-0.01720.36614.348642.9806-8.2229
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 1 through 21 )
2X-RAY DIFFRACTION2chain 'A' and (resid 22 through 47 )
3X-RAY DIFFRACTION3chain 'A' and (resid 48 through 61 )
4X-RAY DIFFRACTION4chain 'A' and (resid 62 through 100 )
5X-RAY DIFFRACTION5chain 'A' and (resid 101 through 116 )
6X-RAY DIFFRACTION6chain 'A' and (resid 117 through 249 )
7X-RAY DIFFRACTION7chain 'A' and (resid 250 through 273 )
8X-RAY DIFFRACTION8chain 'B' and (resid 1 through 82 )
9X-RAY DIFFRACTION9chain 'B' and (resid 83 through 100 )
10X-RAY DIFFRACTION10chain 'B' and (resid 101 through 198 )
11X-RAY DIFFRACTION11chain 'B' and (resid 199 through 273 )
12X-RAY DIFFRACTION12chain 'C' and (resid 5 through 39 )
13X-RAY DIFFRACTION13chain 'C' and (resid 40 through 61 )
14X-RAY DIFFRACTION14chain 'C' and (resid 62 through 115 )
15X-RAY DIFFRACTION15chain 'C' and (resid 116 through 249 )
16X-RAY DIFFRACTION16chain 'C' and (resid 250 through 273 )
17X-RAY DIFFRACTION17chain 'D' and (resid 5 through 21 )
18X-RAY DIFFRACTION18chain 'D' and (resid 22 through 53 )
19X-RAY DIFFRACTION19chain 'D' and (resid 54 through 100 )
20X-RAY DIFFRACTION20chain 'D' and (resid 101 through 116 )
21X-RAY DIFFRACTION21chain 'D' and (resid 117 through 176 )
22X-RAY DIFFRACTION22chain 'D' and (resid 177 through 249 )
23X-RAY DIFFRACTION23chain 'D' and (resid 250 through 273 )
24X-RAY DIFFRACTION24chain 'E' and (resid 1 through 48 )
25X-RAY DIFFRACTION25chain 'E' and (resid 49 through 82 )
26X-RAY DIFFRACTION26chain 'E' and (resid 83 through 249 )
27X-RAY DIFFRACTION27chain 'E' and (resid 250 through 273 )

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