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2.4.2.7: adenine phosphoribosyltransferase

This is an abbreviated version!
For detailed information about adenine phosphoribosyltransferase, go to the full flat file.

Word Map on EC 2.4.2.7

Reaction

AMP
+
diphosphate
=
adenine
 +
5-phospho-alpha-D-ribose 1-diphosphate

Synonyms

adenine phosphoribosyl transferase, adenine phosphoribosylpyrophosphate transferase, adenine phosphoribosyltransferase, adenine phosphoribosyltransferase 2, adenione PRTase, adenosine phosphoribosyltransferase, adenylate pyrophosphorylase, adenylic pyrophosphorylase, AMP pyrophosphorylase, AMP-pyrophosphate phosphoribosyltransferase, AMP:pyrophosphate phospho-D-ribosyltransferase, APRT, APRT1, APRT2, APRTase, APRTh, apT, APT1, APT2, APT4, APT5, ApTR, FtAPRT, gpT-1, hAPRT, More, phosphoribosyltransferase, adenine, ScAPRT, SmAPRT1, Smp_054360, SSO2342, transphosphoribosidase, TtAPRT2, TTC1249, TTC1250, TTH1250, TTHA1614, TthAPRT, TthHB27APRT, TT_C1249, TT_C1250, type I PRT

ECTree

     2 Transferases
         2.4 Glycosyltransferases
             2.4.2 Pentosyltransferases
                2.4.2.7 adenine phosphoribosyltransferase

Crystallization

Crystallization on EC 2.4.2.7 - adenine phosphoribosyltransferase

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme in apoform and in complex with substrate adenine, sitting drop vapor diffusion method, mixing of 10 mg/ml protein in 20 mM HEPES, pH 7.5, 150 mM NaCl, 1 mM DTT, and 10% glycerol, with crystallization solutions (a) 0.1 M Tris/HCl pH 8.5, 30% PEG 4000 and 0.2 M MgCl2 and (b) 0.2 M sodium acetate trihydrate, 0.1 M Tris-HCl, pH 8.5, 30% PEG 4000, X-ray diffraction structure determination and analysis at 1.9-2.28 A resolution
enzyme, 10 mg/ml, in complex with 9-deazaadenine and sulfate or Mg-phosphoribosyldiphosphate, 50 mM Hepes, pH 6.0, 8 mM MgCl2, 1 mM DTT, 1:2 molar ratio of 9-deazaadenine and iminoribitol, 1 mM sodium diphosphate, after 45 min incubation preparation of crystallization drops, crystals are obtained from mother liquid 0.1 M sodium acetate, pH 4.6, 24% polyethylene glycol 4000, 0.2 M ammonium sulfate, 0.05 M urea, 18°C, X-ray diffraction structure analysis, hydrogen bond network in the complexes
comparison of the crystal structure of PRPP-Mg2+-bound hAPRT to the ADE/PRPP-Mg2+ and AMP complex structures
enzyme complexes phosphate-hAPRT, hypoxanthine-PRPP-Mg2+-hAPRT, IMP-hAPRT, and GMP-hAPRT, mixing of 400 nl of 5 mg/ml protein complexes in 20 mM Tris-HCl, pH 7.4, 5 mM MgCl2, with 200 nl of crystallization solution made of 85 mM Tris-HCl, pH 8.5, 170 mM NaOAc, 19-21% PEG 4000, and 0-30% glycerol, overnight at 20°C, X-ray diffraction structrue determination and analysis at resolution 1.55-1.90 A, molecular replacement using the structure with PDB ID 6FCH as template, and modeling
hanging drop vapour diffusion method, with 15% (v/v) glycerol, 25.5% (w/v) PEG 4000, 0.17 M sodium acetate, and 0.085 M Tris-HCl, pH 8.5
recombinant human APRT is crystallized in complex with adenosine 5'-monophosphate, hanging-drop vapour-diffusion method
vapour diffusion method, hanging drops from solution: 10 mg/ml purified apo-enzyme in 10 mM MES, pH 6.0, 1 mM dithiothreitol, 5 mM MgCl2, 4°C, reservoir solution: 7-11% polyethylene glycol 5000 monomethyl ether, 0.2 M ammonium acetate, 0.1 M sodium citrate, pH 4.9, 10 mM MgCl2, 1.2-1.6 M ammonium sulfate, for AMP- or adenine-bound crystals addition of 10 mM AMP or 5 mM adenine in the reservoir solution, structure analysis
enzyme in complex with adenosine-5'-monophosphate and a phosphate ion, crystallization at 4°C by hanging-drop vapor-diffusion method
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four crystal structures: (1) a structure (the enzyme/Pi complex) refined at 2.4 A with inorganic phosphate or sulfate bound in the 5-phosphoribosyl binding pocket, (2) an adenine bound structure (the enzyme/adenine complex) refined at 2.4 A, which shows adenine together with phosphates both at the 5'-phosphoryl and PPi positions of the presumed PRPP binding site, (3) an AMP bound structure (the enzyme/AMP complex) refined at 2.4 A, and (4) an ADP bound structure (the enzyme/ADP complex), refined at 2.8 A containing the inhibitor ADP bound like AMP with both the alpha- and beta-phosphates occupying the 5'-phosphoribosyl binding site. No crystals of the enzyme in complex with 5-phosphoribosyl-alpha-1-pyrophosphate are obtained, likely because the enzyme catalyzes a slow breakdown of 5-phosphoribosyl-alpha-1-pyrophosphate to ribose 5-phosphate and PPi. The crystal structure suggests that the enzyme evolves from a 6-oxopurine phosphoribosyltransferase. The individual subunit adopts an overall structure that resembles a 6-oxopurine phosphoribosyltransferase (PRTase) more than known adenine phosphoribosyltransferases implying that adenine phosphoribosyltransferase functionality in Crenarchaeotae has its evolutionary origin in this family of 6-oxopurine phosphoribosyltransferases. The N-terminal two-thirds of the polypeptide chain folds as a traditional type I PRTase with a five-stranded beta-sheet surrounded by helices. The C-terminal third adopts an unusual three-helix bundle structure that together with the nucleobase-binding loop undergoes a conformational change upon binding of adenine and phosphate resulting in a slight contraction of the active site
mixing of protein solution 13-15 mg/ml with an equal volume of mother liquid 0.1 M Hepes, pH 7.5, 1.5 M lithium sulfate, then equilibration against mother liquid at 18°C, crystals appear after 3 days, X-ray diffraction structure analysis, also crystallization of the enzyme in presence of diphosphate, Mg2+ or inhibitor immucillin, which do not bind at the active site
-
purified His-tagged recombinant enzyme in complex with inhibitors D-DIAB and L-DIAB, and also with adenine, X-ray diffraction structure determination and analysis of enzyme-inhibitor complexes at 1.78 A and 1.98 A resolution, respectively, modeling, structure comparisons
purified recombinant enzyme APRT2, hanging-drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 200 mM NaCl and 20 mM Tris-HCl, pH 8.0, with 0.001 ml of precipitant solution containing 19% PEG 20000 and 0.1 M sodium citrate, pH 6.0, 30-50 days, 20°C, X-ray diffraction structure determination and analysis at 2.6 A resolution, molecular replacement using the structure of TthAPRT1 from Thermus thermophilus HB8 (PDB ID 1VCH) as a template
purified recombinant enzyme, crystals grow via capillary counter-diffusion technique, X-ray diffraction structure determination and analysis at 2.5 A resolution, molecular replacement amd modeling
purified recombinant enzyme, hanging drop vapor diffusion technique, mixing of 12.5 mg/ml protein in 0.02 M Tris-HCl buffer, pH 8.0, 50 mM NaCl, 5% glycerol, 0.04% NaN3, and 5 mM AMP, with reservoir solution composed of 25% w/v PEG 3350, 0.1 M HEPES, pH 7.0, 0.5 M NaCl, and 0.04% NaN3, X-ray diffraction structure determination and analysis at 2.60 A resolution
purified recombinant native and selenomethionine-labeled enzymes, sitting drop vapour diffusion method, 0.001 ml of 20 mg/ml native protein in 20 mM Tris-HCl, 50 mM NaCl, pH 8.0, is mixed with an equal volume of reservoir solution containing 100 mM MES, pH 5.5, 100 mM calcium acetate, 3% w/v PEG 10000, and 3% v/v MeOH, 1 week, for the selenomethionine-labeled protein 40 mM calcium acetate, 1.5% w/v PEG 10000, and 3% v/v MeOH is used at a protein concentration of 29.4 mg/ml, X-ray diffraction structure determination and analysis at 1.9-2.6 A resolution, multiple wavelength anomalous dispersion method, asymmetric unit of two pairs of identical dimers, each related by noncrystallographic two-fold symmetry, a fifth monomer forms a similar dimer across a crystallographic two-fold axis, modeling
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