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2.4.2.8: hypoxanthine phosphoribosyltransferase

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

Word Map on EC 2.4.2.8

Reaction

IMP
+
diphosphate
=
hypoxanthine
+
5-phospho-alpha-D-ribose 1-diphosphate

Synonyms

6-hydroxypurine phosphoribosyltransferase, 6-mercaptopurine phosphoribosyltransferase, 6-oxo PRTase, 6-oxopurine phosphoribosyltransferase, 6-oxopurine PRTase, Cthe_2254, EcHPRT, GMP pyrophosphorylase, GPRT, gpT-2, guanine phosphoribosyltransferase, guanine PRTase, guanine-hypoxanthine phosphoribosyltransferase, guanosine 5'-phosphate pyrophosphorylase, guanosine phosphoribosyltransferase, guanylate pyrophosphorylase, guanylic pyrophosphorylase, HGPRT, HGPRT-1, HGPRTase, HGXPRT, HGXPRTase, HG[X]PRT, HPRT, HPRT1, HPRTJerusalem, HPT, HXGPRT, hypoxanthine guanine phosphoribosyltransferase, hypoxanthine guanine xanthine phosphoribosyltransferase, hypoxanthine phosphoribosyl transferase, hypoxanthine phosphoribosyltransferase, hypoxanthine phosphoribosyltransferase 1, hypoxanthine PRTase, hypoxanthine-guanine phosphoribosyl transferase, hypoxanthine-guanine phosphoribosyltransferase, hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase, hypoxanthine-guanine-(xanthine) phosphoribosyltransferase, hypoxanthine-guanine-xanthine phosphoribosyltransferase, hypoxanthine-guanine-[xanthine]-phosphoribosyltransferase, hypoxanthine-xanthine-guanine phosphoribosyltransferase, IMP pyrophosphorylase, IMP-GMP pyrophosphorylase, inosinate pyrophosphorylase, inosine 5'-phosphate pyrophosphorylase, inosine monophosphate:pyrophosphate phosphoribosyltransferase, inosinic acid pyrophosphorylase, inosinic pyrophosphorylase, More, MSMEG_6110, MtHGPRT, Pf HGXPRT, PF3D7_1012400, PfHGXPRT, PhHGXPRT, phosphoribosyltransferase, 6-mercaptopurine, phosphoribosyltransferase, hypoxanthine, PhPRT, plasma hypoxanthine-guanine phosphoribosyl transferase, PRT, purine-6-thiol phosphoribosyltransferase, PvHGPRT, SmHGPRT1, SSO2424, Tb927.10.1400, TcHPRT, transphosphoribosidase, TthHPRT, TT_C1766, ZgHGPRT/AMPK

ECTree

     2 Transferases
         2.4 Glycosyltransferases
             2.4.2 Pentosyltransferases
                2.4.2.8 hypoxanthine phosphoribosyltransferase

Crystallization

Crystallization on EC 2.4.2.8 - hypoxanthine phosphoribosyltransferase

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
free enzyme or enzyme complexed with IMP, hanging drop vapour diffusion method, 20°C, 10 mg/ml protein in 20 Tris-HCl, pH 7.4, mixed with a molar excess of IMP, 0.001 ml protein solution mixed with equal volume of reservoir solution containing 0.2 M magnesium acetate tetrahydrate, 0.1 M sodium cacodylate, pH 6.5, 12% w/v PEG 4000, or protein solution with 1 mM IMP mixed with reservoir solution containing 15% PEG 8000, 0.1 M sodium cacodylate, pH 6.5, 0.2 M magnesium acetate tetrahydrate, X-ray diffraction structure determination and analysis at 2.2-2.5 A resolution
-
purified recombinant mutant L160I enzyme, hanging drop vapour diffusion method, 0.001 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH 7.4, excess GMP in a molar ratio 4:1, is mixed with equal volume of crystallization solution containing 0.2 M CaCl2 dihydrate, 0.1 M HEPES, pH 7.5, 28% PEG 400, 20°C, 2 weeks, X-ray diffraction structure determination and analysis at 1.7 A resolution, purified recombinant wild-type enzyme, hanging drop vapour diffusion method, 0.001 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH 7.4, excess IMP in a molar ratio 4:1, is mixed with equal volume of crystallization solution containing 0.2 M magnesium acetate tetrahydrate, 0.1 M sodium cacodylate, pH 6.5, 12% PEG 4000, 20°C, within 1 week, X-ray diffraction structure determination and analysis at 2.2 A resolution
-
enzyme in complex with inhibitors (2-[(2,3-dihydroxypropyl)[2-(6-oxo-1,6-dihydro-9H-purin-9-yl)ethyl]amino]ethyl)phosphonic acid, 9-[(N-phosphonoethyl-N-phosphonomethyl)-2-aminoethyl]-hypoxanthine, 9-[(N-phosphonoethyl-N-phosphonoethoxyethyl)-2-aminoethyl]-hypoxanthine, (2-[[2-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)ethyl][2-(2-phosphonoethoxy)ethyl]amino]ethyl)phosphonic acid, [[2-[(6-oxo-1,6-dihydro-9H-purin-9-yl)methyl]propane-1,3-diyl]bis(oxymethylene)]bis(phosphonic acid), and [[[(4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-7-yl)methyl]azanediyl]di(propane-3,1-diyl)]bis(phosphonic acid), X-ray diffraction structure determination and analysis at 2.55-2.9 A resolution
enzyme in complex with 2-(phosphonoethoxy)ethyl guanine, 2-(phosphonoethoxy)ethyl hypoxanthine, and (R,S)-9-[3-hydroxy-2-(phosphonomethoxy)propyl]guanine, hanging drop vapor diffusion method, mixing of equal volumes of well solution, containing 0.1 M citrate pH 5.5, 10% isopropyl alcohol and 29% PEG 4000, and protein inhibitor complex with 17 mg/ml prtoein, and 3.3 mM for 2-(phosphonoethoxy)ethyl guanine, 3.9 mM (R,S)-9-[3-hydroxy-2-(phosphonomethoxy)propyl]guanine, and 3.0 mM for 2-(phosphonoethoxy)ethyl hypoxanthine, X-ray diffraction structure determination and analysis at 2.6-2.78 A resolution
in complex with T-705-RMP, hanging drop vapor diffusion method, using 0.2 M NaCl, 1 M sodium/potassium tartrate, 0.1 M imidazole, pH 8.0
purified enzyme in complex with inhibitor [3-(guanine-9-yl)-2-((2-phosphonoethoxy)methyl)propoxy]methylphosphonic acid and ([3-(guanine-9-yl)-2-((2-phosphonoethoxy)-methyl)propoxy]methyl)phosphonic acid, hanging drop method, mixing of 0.001 ml of 11.1 mg/ml protein in 0.1 M Tris-HCl, 0.01 M MgCl2, 1 mM DTT, and 0.3 mM 5-phospho-alpha-D-ribose 1-diphosphate, pH 7.4, and 4.8 mM inhibitor, with 0.001 ml of reservoir solution containing 20% PEG 3350, 0.2 M sodium bromide, 0.1 M Bis-Tris propane, pH 7.5, at 18°C, X-ray diffraction structure determination and analysis at 2.0 A resolution, molecular replacement and modeling
purified recombinant mutant C22A/C105A/C205A enzyme 1. free or 2. in complex with inactive purine base analogue 7-hydroxy [4,3d] pyrazolo pyrimidine and 5-phospho-alpha-D-ribose 1-diphosphate, or 3. complexed with IMP or GMP, or 4. complexed with transition state analogue immuncillinHP-Mg2+-diphosphate, hanging drop vapour diffusion method, 18 mg/ml protein in 0.05 M Tris-HCl, pH 7.4, 1 mM MgCl2, 1 mM DTT, mixing of 0.002 ml of both protein and reservoir solution, the latter containing 0.2 M ammonium acetate, 0.1 m sodium acetate, pH 4.6, 30% w/v PEG 4000, 17°C, 2-7 days, cryoprotection by 30% glycerol in reservoir solution, X-ray diffraction structure determination and analysis at 1.9 A resolution
-
recombinant chimeric mutant enzyme complex with the product GMP, 12 mg/ml protein and 5 mM GMP in 0.1 M Tris, pH 8.0, and 2.0 M ammonium sulfate, 2-5 days, X-ray diffraction structure determination and analysis at 2.8 A resolution, modeling
spectral analysis of the crystal structure of the HGPRT/immucillin-G 5'-phosphate/diphosphate complex
ultraviolet resonance Raman spectroscopy study on the complexes of enzyme with products IMP, GMP, and XMP, both in Homo sapiens and Plasmodium falciparum, in resonance with the purine nucleobase electronic absorption. Human hypoxanthine guanine phosphoribosyltransferase catalyzes the phosphoribosylation of guanine and hypoxanthine, while the Plasmodium falciparum enzyme acts on xanthine as well. Spectra of bound nucleotides show that the enzyme distorts the structure of the nucleotides. The distorted structure resembles that of the deprotonated nucleotide. The two proteins assemble similar active sites for their common substrates. While the human enzyme does not bind XMP, Plasmodium falciparum hypoxanthine guanine phosphoribosyltransferase perturbs the pKa of bound XMP
apo-form and in complex with GMP, sitting drop vapor diffusion method, using 0.2 M trimethylamine N-oxide dehydrate, 0.1 M Tris pH 8.5, 20% (w/v) polyethylene glycol monomethyl ether 2000
recombinant enzyme, 7 mg/ml, hanging-drop vapour-diffusion method, TMD buffer, pH 7.5, + equal volume of reservoir solution: 18°C or 4°C, pH 5.6 , 19% isopropanol, 19% polyethylene glycol 4000, 5% glycerol, or 17% polyethylene glycol 4000, 5% glycerol
purified recombinant enzyme PvHGPRT in complex with inhibitor [3R,4R]-4-guanin-9-yl-3-((S)-2-hydroxy-2-phosphonoethyl)oxy-1-N-(phosphonopropionyl)pyrrolidine, 21 mg/ml protein is incubated with 6.4 mM inhibitor on ice for 5 min, hanging drop method, mixing of 0.001 ml of protein and inhibitor with 0.001 ml of reservoir solution containing 0.3 M potassium thiocyanate, 0.1 M Bis-tris propane, pH 8.5, 18°C, X-ray diffraction structure determination and analysis at 2.85 A resolution, molecular replacement using the protein coordinates of subunit A of human HGPRT in complex with [(2-[(guanin-9H-yl)methyl]-propane-1,3-diyl)bis(oxy)]bis-(methylene)diphosphonic acid as template (PDB ID 4IJQ), model building
recombinant chimeric mutant enzyme complex with the product GMP, 12 mg/ml protein and 5 mM GMP in 0.1 M Tris, pH 8.0, and 2.0 M ammonium sulfate, 2-5 days, X-ray diffraction structure determination and analysis at 2.8 A resolution, modeling
ultraviolet resonance Raman spectroscopy study on the complexes of enzyme with products IMP, GMP, and XMP, both in Homo sapiens and Plasmodium falciparum, in resonance with the purine nucleobase electronic absorption. Human hypoxanthine guanine phosphoribosyltransferase catalyzes the phosphoribosylation of guanine and hypoxanthine, while the Plasmodium falciparum enzyme acts on xanthine as well. Spectra of bound nucleotides show that the enzyme distorts the structure of the nucleotides. The distorted structure resembles that of the deprotonated nucleotide. The two proteins assemble similar active sites for their common substrates. While the human enzyme does not bind XMP, Plasmodium falciparum hypoxanthine guanine phosphoribosyltransferase perturbs the pKa of bound XMP
-
purified isozyme HPGRT-1 in complex with IMP, 2,5 mg/ml enzyme and 2 mM of IMP are mixed with 12.5% PEG 1000, 12.5% PEG 3350, 12.5% MPD, 0.03 M divalent cations, and 0.1 M MES/imidazole, pH 6.5, at 18°C, X-ray diffraction structure determination and analysis at 2.8 A resolution, screening and method optimmization, HPGRT-1 crystallizes in an orthorhombic system, and the systematic absences indicates space group P212121 with four polypeptide chains in the asymmetric unit, respectively
structure in the unliganded form, in complex with IMP and in complex with GMP, to 2.1, 1.9 and 2.2 A resolution, respectively. The overall fold of the IMP complex is similar to that of the unliganded form, but the main-chain and side-chain atoms of the active site move to accommodate IMP. The overall folds of the IMP and GMP complexes are almost identical to each other. Enzyme belongs to group I
crystallization in complex with GMP and IMP, structure analysis
mutant enzyme D150A, crystallization complexed with xanthosine 5'-monophosphate, diphosphate and 2 Mg2+, post transition state structure analysis, active site structure
the recombinant enzyme is complexed with Mg2+, 5-phosphoribosyl 1-diphosphate and inactive substrate analogue 9-deazaguanine, hanging drop method, enzyme complex, 20 mg/ml, is precipitated by 0.1 M Tris-HCl, pH 8.0, 30% polyethylene glycol 4000, 0.2 M Li2SO4, 0.5% beta-octylglucoside at 4°C, X-ray diffraction structure analysis
-
analysis of crystal structure
-
crystallization and X-ray structure determination and analysis, 1 molecule of GMP bound per dimer
-
purified recombinant TbrHGPRT in complex with GMP, IMP, and inhibitors [6-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)hexyl]phosphonic acid, [4-(6-oxo-1,6-dihydro-9H-purin-9-yl)butyl]phosphonic acid, and [5-(6-oxo-1,6-dihydro-9H-purin-9-yl)pentyl]phosphonic acid, hanging drop vapor diffusion method, mixing of 0.001 ml of protein ligand complex solution and 0.001 ml of well solution. The protein-ligand complexes are prepared using 36 mg/ml, 28 mg/ml, 60 mg/ml, and 28 mg/ml of enzyme in the presence of 3.2 mM GMP, 3.2 mM IMP, 4.6 mM [5-(6-oxo-1,6-dihydro-9H-purin-9-yl)pentyl]phosphonic acid, 4.6 mM [4-(6-oxo-1,6-dihydro-9H-purin-9-yl)butyl]phosphonic acid and 1.8 mM [6-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)hexyl]phosphonic acid, respectively, followed by incubation on ice for 10 min, the reservoir solution for the complexes with GMP, IMP, [4-(6-oxo-1,6-dihydro-9H-purin-9-yl)butyl]phosphonic acid, and [6-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)hexyl]phosphonic acid contains 25% PEG 3350, 0.2 M lithium sulfate, and 0.1 M Bis-Tris, pH 5.0-5.5. For [5-(6-oxo-1,6-dihydro-9H-purin-9-yl)pentyl]phosphonic acid the well solution is 25% PEG 3350, 0.2 M sodium iodide, 0.1 M Bis-Tris propane, pH 6.5, X-ray diffraction structure determination and analysis at 2.73, 2.51, 1.52, 2.89 and 2.81 A resolution, respectively. The structure of the complex with [6-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)hexyl]phosphonic acid is solved by molecular replacement using the structure of TcrHGPRT (PDB ID 1TC2) as the starting model. The starting models for the complexes with GMP, IMP, [4-(6-oxo-1,6-dihydro-9H-purin-9-yl)butyl]phosphonic acid, and [5-(6-oxo-1,6-dihydro-9H-purin-9-yl)pentyl]phosphonic acid are based upon the refined model of the complex with [6-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)hexyl]phosphonic acid, refinement and model building
purified recombinant His6-tagged enzyme, mixing of 5 mg/ml protein in 20 mM Tris-HCl, 100 mM NaCl, pH 8.0, with reservoir solution containing 0.1 M MES, pH 6.5, and 12% w/v PEG 20000 at 20°C, X-ray diffraction structure determination and analysis at 2.65 A resolution