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- PDB-1ia3: Candida albicans dihydrofolate reductase complex in which the dih... -
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Open data
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Basic information
Entry | Database: PDB / ID: 1ia3 | ||||||
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Title | Candida albicans dihydrofolate reductase complex in which the dihydronicotinamide moiety of dihydro-nicotinamide-adenine-dinucleotide phosphate (NADPH) is displaced by 5-[(4-TERT-BUTYLPHENYL)SULFANYL]-2,4-QUINAZOLINEDIAMINE (GW995) | ||||||
![]() | DIHYDROFOLATE REDUCTASE | ||||||
![]() | OXIDOREDUCTASE / ANTIFUNGAL TARGET / REDUCTASE / Dihydronicotinamide displaced | ||||||
Function / homology | ![]() dihydrofolate metabolic process / glycine biosynthetic process / dihydrofolate reductase / dihydrofolate reductase activity / folic acid metabolic process / tetrahydrofolate biosynthetic process / one-carbon metabolic process / NADP binding / mitochondrion / cytosol Similarity search - Function | ||||||
Biological species | ![]() | ||||||
Method | ![]() | ||||||
![]() | Whitlow, M. / Howard, A.J. / Kuyper, L.F. | ||||||
![]() | ![]() Title: X-ray Crystal Structures of Candida albicans Dihydrofolate Reductase: High Resolution Ternary Complexes in Which the Dihydronicotinamide Moiety of NADPH is Displaced by an inhibitor Authors: Whitlow, M. / Howard, A.J. / Stewart, D. / Hardman, K.D. / Chan, J.H. / Baccanari, D.P. / Tansik, R.L. / Hong, J.S. / Kuyper, L.F. #1: ![]() Title: X-ray Crystallographic Studies of Candida albicans Dihydrofolate Reductase. High resolution structures of the holoenzyme and an inhibited ternary complex. Authors: Whitlow, M. / Howard, A.J. / Stewart, D. / Hardman, K.D. / Kuyper, L.F. / Baccanari, D.P. / Fling, M.E. / Tansik, R.L. #2: ![]() Title: Selective Inhibitors of Candida albicans Dihydrofolate Reductase: Activity and Selectivity of 5-(arylthio)-2,4-diaminoquinazolines Authors: Chan, J.H. / Hong, J.S. / Kuyper, L.F. / Baccanari, D.P. / Joyner, S.S. / Tansik, R.L. / Boytos, C.M. / Rudolph, S.K. #3: ![]() Title: Characterization of Candida Albicans Dihydrofolate Reductase Authors: Baccanari, D.P. / Tansik, R.L. / Joyner, S.S. / Fling, M.E. / Smith, P.L. / Freisheim, J.H. | ||||||
History |
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Remark 99 | HET GROUPS NDP 193, TQ5 194, AND MES 201 ARE ASSOCIATED WITH PROTEIN CHAIN A. HET GROUPS NDP 195, ...HET GROUPS NDP 193, TQ5 194, AND MES 201 ARE ASSOCIATED WITH PROTEIN CHAIN A. HET GROUPS NDP 195, TQ5 196, AND MES 202 ARE ASSOCIATED WITH PROTEIN CHAIN B. | ||||||
Remark 600 | HETEROGEN THE INHIBITION CONSTANT (IC50) FOR 5-[(4-TERT-BUTYLPHENYL) SULFANYL]-2,4- ... HETEROGEN THE INHIBITION CONSTANT (IC50) FOR 5-[(4-TERT-BUTYLPHENYL) SULFANYL]-2,4-QUINAZOLINEDIAMINE (GW995) IS 8 NANOMOLAR USING THE FOLLOWING ASSAY (SEE REFERENCE 0 & 3). C. ALBICANS DHFR ASSAY WAS PERFORMED IN 0.1 M IMIDAZOLE CHLORIDE BUFFER, PH 6.4, WITH 12 MM MERCAPTOETHANOL, 60 MM NADPH AND 45 MM DIHYDROFOLIC ACID IN A FINAL VOLUME OF 1 ML AT 303K. IC50 IS THE CONCENTRATION OF INHIBITOR THAT DECREASES THE VELOCITY OF THE STANDARD ASSAY BY 50%. THE ENZYME (0.2 NM), NADPH, AND VARYING CONCENTRATIONS OF INHIBITOR WERE PREINCUBATED FOR 2 MIN AT 30-C, AND THE REACTION WAS INITIATED BY THE ADDITION OF DIHYDROFOLIC ACID. STEADY STATE VELOCITIES WERE MEASURED, AND IC50 VALUES WERE CALCULATED FROM A LINEAR REGRESSION PLOT OF THE PERCENTAGE INHIBITION VS THE LOGARITHM OF THE INHIBITOR CONCENTRATION. IC50 THE PRECISION OF THE DETERMINATION IS GENERALLY ABOUT A 30%. |
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Structure visualization
Structure viewer | Molecule: ![]() ![]() |
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Downloads & links
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Download
PDBx/mmCIF format | ![]() | 106.9 KB | Display | ![]() |
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PDB format | ![]() | 80.8 KB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
Others | ![]() |
-Validation report
Summary document | ![]() | 635.1 KB | Display | ![]() |
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Full document | ![]() | 654.4 KB | Display | |
Data in XML | ![]() | 12.9 KB | Display | |
Data in CIF | ![]() | 19.6 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 1ia1C ![]() 1ia2C ![]() 1ia4C ![]() 1ai9S S: Starting model for refinement C: citing same article ( |
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Similar structure data |
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Links
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Assembly
Deposited unit | ![]()
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2 | ![]()
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Unit cell |
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Details | Dihydrofolate reductase is active as a monomer. |
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Components
#1: Protein | Mass: 22194.527 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() #2: Chemical | #3: Chemical | #4: Chemical | #5: Water | ChemComp-HOH / | |
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-Experimental details
-Experiment
Experiment | Method: ![]() |
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Sample preparation
Crystal | Density Matthews: 2.27 Å3/Da / Density % sol: 43 % | ||||||||||||||||||||||||||||||
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Crystal grow | Temperature: 277 K / Method: vapor diffusion, hanging drop / pH: 6.5 Details: DIHYDRO-NICOTINAMIDE-ADENINE- DINUCLEOTIDE PHOSPHATE (NADPH),5-[(4-TERT-BUTYLPHENYL) SULFANYL]-2,4-QUINAZOLINEDIAMINE (GW995), PEG-3350, POTASSIUM 4-MORPHILINEERHANESULFONIC ACID, ...Details: DIHYDRO-NICOTINAMIDE-ADENINE- DINUCLEOTIDE PHOSPHATE (NADPH),5-[(4-TERT-BUTYLPHENYL) SULFANYL]-2,4-QUINAZOLINEDIAMINE (GW995), PEG-3350, POTASSIUM 4-MORPHILINEERHANESULFONIC ACID, DITHIOTHREITOL A THREE-FOLD EXCESS OF GW995 AND THREE-FOLD EXCESS OF NADPH WAS ADDED TO THE C. ALBICANS DHFR SOLUTION AND LET STAND 277K OVERNIGHT. 17-20 MG/ML C. ALBICANS DHFR IN 50 UM GW995, 50 UM NADPH, 20 MM KMES, 1 MM DTT, PH 6.5 WAS MIXED WITH AN EQUAL PART OF 26 - 34% PEG-3350, THE RESERVOIR SOLUTION. , VAPOR DIFFUSION, HANGING DROP | ||||||||||||||||||||||||||||||
Crystal grow | *PLUS Temperature: 4 ℃ | ||||||||||||||||||||||||||||||
Components of the solutions | *PLUS
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-Data collection
Diffraction | Mean temperature: 293 K |
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Diffraction source | Source: ![]() |
Detector | Type: XENTRONICS / Detector: AREA DETECTOR / Date: Aug 30, 1988 / Details: Huber graphite monochromator |
Radiation | Monochromator: Huber graphite monochromator / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray |
Radiation wavelength | Wavelength: 1.5418 Å / Relative weight: 1 |
Reflection | Resolution: 1.75→25 Å / Num. all: 116944 / Num. obs: 36058 / % possible obs: 90.1 % / Observed criterion σ(F): 0 / Observed criterion σ(I): -3 / Redundancy: 3.24 % / Biso Wilson estimate: 36.3 Å2 / Rmerge(I) obs: 0.0549 / Rsym value: 0.0549 / Net I/σ(I): 16.18 |
Reflection shell | Resolution: 1.75→1.85 Å / Redundancy: 1.11 % / Rmerge(I) obs: 0.2694 / Mean I/σ(I) obs: 2.23 / Num. unique all: 4005 / Rsym value: 0.2694 / % possible all: 60.4 |
Reflection shell | *PLUS % possible obs: 60.4 % |
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Processing
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Refinement | Method to determine structure: DIRECT REPLACEMENT Starting model: CANDIDA ALBICANS DHFR (PDB ENTRY 1AI9) Resolution: 1.78→10 Å Isotropic thermal model: Konnert, J.H. & Hendrickson, W.A. (1980) Acta Crystallogr. A A36, 344. σ(F): 2 / σ(I): 0 Stereochemistry target values: Hendrickson, W.A. (1985) Methods Enzymol. 115, 252-270. Details: Restrain least-squares procedure
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Refinement step | Cycle: LAST / Resolution: 1.78→10 Å
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Refine LS restraints |
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LS refinement shell |
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