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- PDB-7l28: Crystal structure of the catalytic domain of human PDE3A bound to... -

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Basic information

Entry
Database: PDB / ID: 7l28
TitleCrystal structure of the catalytic domain of human PDE3A bound to Trequinsin
ComponentscGMP-inhibited 3',5'-cyclic phosphodiesterase A
KeywordsHYDROLASE/HYDROLASE INHIBITOR / HYDROLASE / HYDROLASE-HYDROLASE INHIBITOR complex
Function / homology
Function and homology information


cGMP-inhibited cyclic-nucleotide phosphodiesterase activity / estrogen binding / regulation of ribonuclease activity / positive regulation of oocyte development / regulation of meiotic nuclear division / cellular response to cGMP / oocyte maturation / positive regulation of vascular permeability / negative regulation of vascular permeability / negative regulation of cAMP-mediated signaling ...cGMP-inhibited cyclic-nucleotide phosphodiesterase activity / estrogen binding / regulation of ribonuclease activity / positive regulation of oocyte development / regulation of meiotic nuclear division / cellular response to cGMP / oocyte maturation / positive regulation of vascular permeability / negative regulation of vascular permeability / negative regulation of cAMP-mediated signaling / 3',5'-cyclic-nucleotide phosphodiesterase / nuclear estrogen receptor activity / 3',5'-cyclic-GMP phosphodiesterase activity / cGMP-mediated signaling / cAMP-mediated signaling / 3',5'-cyclic-nucleotide phosphodiesterase activity / 3',5'-cyclic-AMP phosphodiesterase activity / cellular response to transforming growth factor beta stimulus / apoptotic signaling pathway / lipid metabolic process / G alpha (s) signalling events / response to xenobiotic stimulus / G protein-coupled receptor signaling pathway / negative regulation of apoptotic process / membrane / metal ion binding / cytosol
Similarity search - Function
3'5'-cyclic nucleotide phosphodiesterase, catalytic domain / 3'5'-cyclic nucleotide phosphodiesterase, conserved site / 3'5'-cyclic nucleotide phosphodiesterase, catalytic domain superfamily / 3'5'-cyclic nucleotide phosphodiesterase / 3'5'-cyclic nucleotide phosphodiesterase domain signature. / 3'5'-cyclic nucleotide phosphodiesterase domain profile. / Metal dependent phosphohydrolases with conserved 'HD' motif. / HD/PDEase domain
Similarity search - Domain/homology
ACETATE ION / : / Chem-XKG / cGMP-inhibited 3',5'-cyclic phosphodiesterase 3A
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.2 Å
AuthorsHorner, S.W. / Garvie, C.
CitationJournal: Nat Commun / Year: 2021
Title: Structure of PDE3A-SLFN12 complex reveals requirements for activation of SLFN12 RNase.
Authors: Colin W Garvie / Xiaoyun Wu / Malvina Papanastasiou / Sooncheol Lee / James Fuller / Gavin R Schnitzler / Steven W Horner / Andrew Baker / Terry Zhang / James P Mullahoo / Lindsay Westlake / ...Authors: Colin W Garvie / Xiaoyun Wu / Malvina Papanastasiou / Sooncheol Lee / James Fuller / Gavin R Schnitzler / Steven W Horner / Andrew Baker / Terry Zhang / James P Mullahoo / Lindsay Westlake / Stephanie H Hoyt / Marcus Toetzl / Matthew J Ranaghan / Luc de Waal / Joseph McGaunn / Bethany Kaplan / Federica Piccioni / Xiaoping Yang / Martin Lange / Adrian Tersteegen / Donald Raymond / Timothy A Lewis / Steven A Carr / Andrew D Cherniack / Christopher T Lemke / Matthew Meyerson / Heidi Greulich /
Abstract: DNMDP and related compounds, or velcrins, induce complex formation between the phosphodiesterase PDE3A and the SLFN12 protein, leading to a cytotoxic response in cancer cells that express elevated ...DNMDP and related compounds, or velcrins, induce complex formation between the phosphodiesterase PDE3A and the SLFN12 protein, leading to a cytotoxic response in cancer cells that express elevated levels of both proteins. The mechanisms by which velcrins induce complex formation, and how the PDE3A-SLFN12 complex causes cancer cell death, are not fully understood. Here, we show that PDE3A and SLFN12 form a heterotetramer stabilized by binding of DNMDP. Interactions between the C-terminal alpha helix of SLFN12 and residues near the active site of PDE3A are required for complex formation, and are further stabilized by interactions between SLFN12 and DNMDP. Moreover, we demonstrate that SLFN12 is an RNase, that PDE3A binding increases SLFN12 RNase activity, and that SLFN12 RNase activity is required for DNMDP response. This new mechanistic understanding will facilitate development of velcrin compounds into new cancer therapies.
History
DepositionDec 16, 2020Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jun 16, 2021Provider: repository / Type: Initial release
Revision 1.1Jul 28, 2021Group: 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 1.2Oct 18, 2023Group: Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / 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
A: cGMP-inhibited 3',5'-cyclic phosphodiesterase A
B: cGMP-inhibited 3',5'-cyclic phosphodiesterase A
C: cGMP-inhibited 3',5'-cyclic phosphodiesterase A
D: cGMP-inhibited 3',5'-cyclic phosphodiesterase A
hetero molecules


Theoretical massNumber of molelcules
Total (without water)174,85220
Polymers172,6764
Non-polymers2,17516
Water1,928107
1
A: cGMP-inhibited 3',5'-cyclic phosphodiesterase A
B: cGMP-inhibited 3',5'-cyclic phosphodiesterase A
hetero molecules


  • defined by author
  • Evidence: equilibrium centrifugation, Analytical ultracentrifugation suggests a similar PDE3A construct (residues 640-1141) exist as a dimer with a Kd of 40 nM., cross-linking, Cross-linking with NHS ...Evidence: equilibrium centrifugation, Analytical ultracentrifugation suggests a similar PDE3A construct (residues 640-1141) exist as a dimer with a Kd of 40 nM., cross-linking, Cross-linking with NHS and EDC, followed by denaturing SDS-PAGE, yield bands corresponding to 2 times the mass of the monomer.
  • 87.4 kDa, 2 polymers
  • Search similar-shape structures of this assembly by Omokage search (details)
Theoretical massNumber of molelcules
Total (without water)87,42610
Polymers86,3382
Non-polymers1,0888
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
C: cGMP-inhibited 3',5'-cyclic phosphodiesterase A
D: cGMP-inhibited 3',5'-cyclic phosphodiesterase A
hetero molecules


Theoretical massNumber of molelcules
Total (without water)87,42610
Polymers86,3382
Non-polymers1,0888
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)83.240, 59.650, 158.120
Angle α, β, γ (deg.)90.000, 90.480, 90.000
Int Tables number4
Space group name H-MP1211
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
12A
22C
13A
23D
14B
24C
15B
25D
16C
26D

NCS domain segments:

Component-ID: 0 / Beg auth comp-ID: LYS / Beg label comp-ID: LYS / Refine code: 0

Dom-IDEns-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11GLUGLUAA669 - 10932 - 378
21GLUGLUBB669 - 10932 - 378
12GLUGLUAA669 - 10932 - 378
22HISHISCC669 - 10782 - 363
13GLUGLUAA669 - 10932 - 378
23HISHISDD669 - 10782 - 363
14GLNGLNBB669 - 10942 - 379
24LEULEUCC669 - 10792 - 364
15GLNGLNBB669 - 10942 - 379
25LEULEUDD669 - 10792 - 364
16LEULEUCC669 - 10792 - 364
26LEULEUDD669 - 10792 - 364

NCS ensembles :
ID
1
2
3
4
5
6

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Components

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Protein , 1 types, 4 molecules ABCD

#1: Protein
cGMP-inhibited 3',5'-cyclic phosphodiesterase A / Cyclic GMP-inhibited phosphodiesterase A / CGI-PDE A


Mass: 43169.121 Da / Num. of mol.: 4 / Fragment: Catalytic domain
Source method: isolated from a genetically manipulated source
Details: N-terminal glycine from cleavage with TEV protease. Residues 780 to 800 replaced with GGSGGS linker. Residues 1029 to 1067 replaced with GGSGGS linker.
Source: (gene. exp.) Homo sapiens (human) / Gene: PDE3A / Plasmid: pET21 / Production host: Escherichia coli BL21(DE3) (bacteria)
References: UniProt: Q14432, 3',5'-cyclic-nucleotide phosphodiesterase

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Non-polymers , 5 types, 123 molecules

#2: Chemical
ChemComp-XKG / (2E)-9,10-dimethoxy-3-methyl-2-[(2,4,6-trimethylphenyl)imino]-2,3,6,7-tetrahydro-4H-pyrimido[6,1-a]isoquinolin-4-one / Trequinsin / Trequinsin


Mass: 405.489 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C24H27N3O3 / Feature type: SUBJECT OF INVESTIGATION / Comment: inhibitor*YM
#3: Chemical
ChemComp-MN / MANGANESE (II) ION


Mass: 54.938 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Mn
#4: Chemical
ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Mg
#5: Chemical
ChemComp-ACT / ACETATE ION / Acetate


Mass: 59.044 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C2H3O2
#6: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 107 / Source method: isolated from a natural source / Formula: H2O

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Details

Has ligand of interestY

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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: 2.27 Å3/Da / Density % sol: 45.91 %
Crystal growTemperature: 293.15 K / Method: vapor diffusion, hanging drop / pH: 5.9 / Details: MES, PEG 3350, calcium acetate / PH range: 5.7 - 6.1

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: ESRF / Beamline: ID30B / Wavelength: 0.976251 Å
DetectorType: DECTRIS PILATUS3 S 6M / Detector: PIXEL / Date: Jul 26, 2018
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.976251 Å / Relative weight: 1
ReflectionResolution: 2.2→46.29 Å / Num. obs: 77869 / % possible obs: 98.2 % / Redundancy: 3.293 % / Biso Wilson estimate: 55.784 Å2 / CC1/2: 0.997 / Rmerge(I) obs: 0.045 / Rrim(I) all: 0.054 / Χ2: 1.003 / Net I/σ(I): 14.54 / Num. measured all: 256435 / Scaling rejects: 147
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. measured obsNum. possibleNum. unique obsCC1/2Rrim(I) all% possible all
2.2-2.343.3620.4792.394234313250125930.9230.5795
2.34-2.53.3730.2793.993948911813117070.9760.33299.1
2.5-2.73.30.1766.223640411095110320.9830.21199.4
2.7-2.953.3620.1059.9733280995798980.9920.12599.4
2.95-3.33.3190.05915.9230947940993250.9960.07199.1
3.3-3.813.2760.03625.626566821181100.9980.04398.8
3.81-4.663.2350.02735.6222163699568500.9980.03297.9
4.66-6.563.1070.02635.7916515544353150.9970.03297.6
6.56-46.292.8720.03332.228728314630390.9960.04196.6

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Processing

Software
NameVersionClassification
XDSdata reduction
XSCALEdata scaling
REFMAC5.8.0267refinement
PDB_EXTRACT3.27data extraction
PHENIXphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 1SO2

1so2


Resolution: 2.2→46.29 Å / Cor.coef. Fo:Fc: 0.949 / Cor.coef. Fo:Fc free: 0.934 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 0.364 / ESU R Free: 0.24 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS U VALUES : REFINED INDIVIDUALLY
RfactorNum. reflection% reflectionSelection details
Rfree0.2634 3894 5 %RANDOM
Rwork0.2333 ---
obs0.2348 73974 98.69 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso max: 147.79 Å2 / Biso mean: 59.439 Å2 / Biso min: 30 Å2
Baniso -1Baniso -2Baniso -3
1-0.6 Å2-0 Å20.17 Å2
2--4.71 Å20 Å2
3----5.31 Å2
Refinement stepCycle: final / Resolution: 2.2→46.29 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms11944 0 116 107 12167
Biso mean--56.65 44.13 -
Num. residues----1480
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0130.01312372
X-RAY DIFFRACTIONr_bond_other_d0.0350.01711479
X-RAY DIFFRACTIONr_angle_refined_deg1.7121.64816809
X-RAY DIFFRACTIONr_angle_other_deg2.291.58826393
X-RAY DIFFRACTIONr_dihedral_angle_1_deg4.90951467
X-RAY DIFFRACTIONr_dihedral_angle_2_deg34.50223.283661
X-RAY DIFFRACTIONr_dihedral_angle_3_deg14.074152027
X-RAY DIFFRACTIONr_dihedral_angle_4_deg18.9691554
X-RAY DIFFRACTIONr_chiral_restr0.1410.21554
X-RAY DIFFRACTIONr_gen_planes_refined0.0050.0214022
X-RAY DIFFRACTIONr_gen_planes_other0.0070.022898
Refine LS restraints NCS

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

Ens-IDDom-IDAuth asym-IDNumberRms dev position (Å)
11A120160.12
12B120160.12
21A119530.12
22C119530.12
31A120340.11
32D120340.11
41B119940.13
42C119940.13
51B121180.12
52D121180.12
61C121180.12
62D121180.12
LS refinement shellResolution: 2.204→2.261 Å / Rfactor Rfree error: 0 / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.383 278 -
Rwork0.374 5278 -
all-5556 -
obs--96.86 %

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