Abstract
The structure of human HGPRT bound to the transition-state analog immucillinGP and Mg2+-pyrophosphate has been determined to 2.0 Å resolution. ImmucillinGP was designed as a stable analog with the stereoelectronic features of the transition state. Bound inhibitor at the catalytic site indicates that the oxocarbenium ion of the transition state is stabilized by neighboring-group participation from MgPPi and O5'. A short hydrogen bond forms between Asp 137 and the purine ring analog. Two Mg2+ ions sandwich the pyrophosphate and contact both hydroxyls of the ribosyl analog. The transition-state analog is shielded from bulk solvent by a catalytic loop that moves ~25 Å to cover the active site and becomes an ordered antiparallel β-sheet.
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References
Krenitsky, T.A., Papaioannou, R. & Elion, G.B. Human hypoxanthine phosphoribosyl transferase: purification, properties and specificity. J. Biol. Chem. 244, 1263–1270 (1969).
Ullman, B. & Carter, D. Hypoxanthine-guanine phosphoribosyl-transferase as a therapeutic target in protozoan infections. Infect. Agents Dis. 4, 29–40 (1995).
Musick, D.L. Structural features of the phosphoribosyltransferases and their relationship to the human deficiency disorders of purine and pyrimidine metabolism. CRC Crit. Rev. Biochem. 11, 1–34 (1981).
Eads, J.C., Scapin, G., Xu, Y., Grubmeyer, C. & Sacchettini, J.C. The crystal structure of human hypoxanthine-guanine phosphoribosyl-transferase with bound GMP. Cell 78, 325–334 (1994).
Scapin, G., Ozturk, D.H., Grubmeyer, C. & Sacchettini, J.C. The crystal structure of the orotate phosphoribosyltransferase complexed with orotate and α-D-phosphoribosyl-1-pyrophosphate. Biochemistry 34, 10744–10754 (1995).
Somoza, J.R., Chin, M.S., Foda, P.J., Wang C.C. & Fletterick R.J. Crystal structure of the hypoxanthine-guanine-xanthine phosphoribosyl-transferase from the protozoan parasite Tritrichomonas foetus. Biochemistry 35, 7032– 7040 (1996).
Schumacher, M.A., Carter, D., Roos, D.S., Ullman, B. & Brennan R.G. Crystal structures of Toxoplasma gondii HGXPRTase reveal the catalytic role of a long flexible loop. Nature Struct. Biol. 3, 881–887 (1996).
Focia, P.J., Craig, S.P. & Eakin, A.E. Approaching the transition state in the crystal structure of a phosphoribosyltransferase. Biochemistry 37, 17120–17127 (1998).
Krahn, J.M., Kim, J.H., Burns, M.R., Parry, R.J. Zalkin, H. & Smith, J.L. Coupled formation of an amidotransferase interdomain ammonia channel and a phosphoribosyltransferase active site. Biochemistry 36, 11061– 11068 (1997).
Xu, Y., Eads, J., Sacchettini, J.C. & Grubmeyer, C. Kinetic mechanism of human hypoxanthine-guanine phosphoribosyltransferase: rapid phosphoribosyl transfer chemistry. Biochemistry 36, 3700–3712 (1997).
Li, C.M., Tyler, P.C., Furneaux, R.H., Kicska, G., Xu, Y, Grubmeyer, C., Girvin, M. & Schramm, V.L. Transition-state analogues as inhibitors of human and malarial hypoxanthine-guanine phosphoribosyltransferases. Nature Struct. Biol. 6, 582–587 (1999).
Xu, Y. & Grubmeyer, C. Catalysis in human hypoxanthine-guanine phosphoribosyltransferase: Asp-137 acts as a general acid/base. Biochemistry 37, 4114–4124 (1998).
Tao, W., Grubmeyer, C. & Blanchard, J.S. Transition state structure of Salmonella typhimurium orotate phosphoribosyltransferase. Biochemistry 35, 14–21 (1996).
Horenstein, B.A. & Schramm, V.L. Correlation of the molecular electrostatic potential surface of an enigmatic transition state with novel transition state inhibitors. Biochemistry 32, 9917–9925 (1993).
Degano, M., Almo, S.C., Sacchettini, J.C. & Schramm, V.L. Trypanosomal nucleoside hydrolase. A novel mechanism from the structure with a transition-state inhibitor. Biochemistry 37, 6277–6285 (1998).
Berti, P.J. & Schramm, V.L. Transition state structure of the solvolytic hydrolysis of NAD+. J. Am. Chem. Soc. 119, 12069–12078 (1997).
Vos, S., Parry, R.J., Burns, M.R., de Jersey, J. & Martin, J.L. Structures of free and complexed forms of Escherichia coli xanthine-guanine phosphoribosyltransferase. J. Mol. Biol. 282, 875–889 (1998).
Jardim, A. & Ullman B. The conserved serine-tyrosine dipeptide in Leishmania donovani hypoxanthine-guanine phosphoribosyltransferase is essential for catalytic activity. J. Biol. Chem. 272, 8967–8973 (1997).
Hove-Jensen, B., Harlow, K.W., King, C.J. & Switzer, R.L. Phosphoribosylpyrophosphate synthetase of Escherichia coli: properties of the purified enzyme and primary structure of the prs gene. J. Biol. Chem. 267, 6765–6771 (1986).
Wilson, J.M., Stout, J.T., Palella, T.D., Davidson, B.L., Kelley, W.N. & Caskey, C.T. A molecular survey of hypoxanthine-guanine phosphoribosyltransferase deficiency in man. J. Clin. Invest. 77, 188–195 (1986).
Laskowski, R.A., MacArthur, M.W. & Thorton, J.M. PROCHECK: a program to check the stereochemical quality of protein structures. J. Appl. Crystallogr. 26, 283–291 (1993).
Otwinowski, Z. & Minor, W. Processing of x-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).
Navaza, J. AmoRe: an automated package for molecular replacement. Acta Crystallogr. A 50, 157–163 (1994).
Brüunger, A.T. X-PLOR version 3.1. (Yale University Press, New Haven, Connecticut; 1992).
Jones T.A. Interactive computer program graphics: Frodo. Methods Enzymol. B 115, 157–171 (1985).
Johnson, G.G., Eisenberg, L.R. & Migeon, B.R. Human and mouse hypoxanthine guanine phosphoribosyl transferase: dimers and tetramers. Science 203, 174–176 (1979).
Kraulis, P.J. MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures. J. Appl. Crystallogr. 24, 946–950 (1991).
Evan, S.V. SETOR: hardware lighted three dimensional solid model representation of macromolecules. J. Mol. Graphics 11, 134– 138 (1993).
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Shi, W., Li, C., Tyler, P. et al. The 2.0 Å structure of human hypoxanthine-guanine phosphoribosyltransferase in complex with a transition-state analog inhibitor. Nat Struct Mol Biol 6, 588–593 (1999). https://doi.org/10.1038/9376
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DOI: https://doi.org/10.1038/9376
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