PON1 Structure

  • Denis Josse
  • Patrick Masson
  • Cynthia Bartels
  • Oksana Lockridge


Paraoxonase (PON1) is an HDL-bound glycoprotein exhibiting calcium-dependent hydrolase and antioxidative activities, both related to distinct structural domains. Few structural data on PON1 and other related proteins, PON2 and 3, are currently available. The sequence of the 354 amino acid mature enzyme has been deduced from its cDNA. PON1 has 3 Cys residues: Cys-42 and -353 form a disulfide bond, Cys-284 is free. The PON1 hydrophobic N-terminus, predicted as an α-helix, is involved in the association of PON1 with phospholipids. The native enzyme purified from human plasma is not exclusively monomelic. The oligomeric state and size of human PON1 bound to non-ionic detergent molecules depends on the concentration of detergent. Five His (H115, H134, H155, H243, H285), 1 Trp (W281), 2 G1u (E53, E195) and 6 Asp (D54, D169, D183, D269, D279) residues are essential for human PON1 arylesterase and organophosphatase activities. The residues in position 192 and 284, although not essential for the PON1 hydrolase activity, could be close to the active site. PON1 shares common structural and functional features with other lactonohydrolases, mainly PON3. The PON1 3D structure will be established in the near future. It will help to relate PON1 structural and functional properties.

Key Words

paraoxonase organophosphate lactonohydrolase calcium-binding site-directed mutagenesis 


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  1. Adkins S, Gan KN, Mody M, La Du BN (1993) Molecular basis for the polymorphic forms of human serum paraoxonase/arylesterase: glutamine or arginine at position 191, for the respective A or B allozymes. Am Hum Genet 52: 598–608Google Scholar
  2. Aviram M (1999) Does paraoxonase play a role in susceptibility to cardiovascular disease? Mol. Med Today 5: 381–386CrossRefGoogle Scholar
  3. Aviram M, Billecke S, Sorenson R, Bisgaier C, Newton R, Rosenblat M, Erogul J, Hsu C, Dunlop C, La Du B (1998) Paraoxonase active site required for protection against LDL oxidation involves its free sulfhydryl group and is different from that required for its arylesterase/paraoxonase activities: selective action of human paraoxonase allozymes Q and R. Arterioscler Thromb Vasc Biol 18: 1617–1624CrossRefGoogle Scholar
  4. Aviram M, Hardak E, Vaya J, Mahmood S, Milo S, Hoffman A, Billicke S, Draganov D, Rosenblat M (2000) Human serum paraoxonases (PON1) Q and R selectively decrease lipid peroxides in human coronary and carotid atherosclerotic lesions: PON1 esterase and peroxidase-like activities. Circulation, 101, 2510–2517.CrossRefGoogle Scholar
  5. Banzon JA, Kuo JM, Fischer DR, Stang PJ, Raushel FM (1995) Histidine-254 is essential for the inactivation of Phosphodiesterase with the alkynyl phosphate esters and diethyl pyrocarbonate. Biochemistry 34: 750–754CrossRefGoogle Scholar
  6. Banzon JA, Kuo JM, Miles BW, Fischer DR, Stang PJ, Raushel FM (1995) Mechanism-based inactivation of phosphotriesterase by reaction of a critical histidine with a ketene intermediate. Biochemistry 34: 743–749CrossRefGoogle Scholar
  7. Blatter MC, James RW, Messmer S, Barja F, Pometta D (1993) Identification of a distinct human high-density lipoprotein subspecies defined by a lipoprotein-associated protein, K-45. Identity of K-45 with paraoxonase. Eur J Biochem 211: 871–879CrossRefGoogle Scholar
  8. Brodersen DE, Nyborg J, Kjeldgaard M (1999) Zinc-binding site of an S100 protein revealed. Two crystal structures of calcium-bound human psoriasin (S100A7) in the zinc-loaded and zinc-free states. Biochemistry 38: 1695–1704CrossRefGoogle Scholar
  9. Brouillette CG, Anantharamaiah GM (1995) Structural models of human apolipoprotein A-I. Biochim Biophys Acta 1256: 103–129CrossRefGoogle Scholar
  10. Bruce C, Davidson W, Kussie P, Lund-Katz S, Phillips MC, Ghosh R, Tall AR (1995) Molecular determinants of plasma cholesteryl ester transfer protein binding to high density lipoproteins. J Biol Chem 270: 11532–11542CrossRefGoogle Scholar
  11. Chabrière E, Viguié N, Baud D, Josse D, Ferrer J-L, Fontecilla-Camps JC, Masson P (2000) Proceedings of the Bioscience Review-US Army Medical Research and Material Commands, June 4-9, Hunt Valley, MD, USA. (in press)Google Scholar
  12. Choi SS, Forster T (1967) Purification of bovine plasma arylesterase. J Dairy Sci 50: 1088–1091CrossRefGoogle Scholar
  13. Choi SS, Forster TL (1967) Triton X-155 as a stabilizer of bovine plasma arylesterase activity. J Dairy Sci 50: 837–839CrossRefGoogle Scholar
  14. Davies H, Richter RJ, Keifer M, Broomfield CA, Sowalla J, Furlong, CE (1996) The effect of the human serum paraoxonase polymorphism is reversed with diazoxon, soman and sarin. Nat Genet 14: 334–336CrossRefGoogle Scholar
  15. Don MM, Masters CJ, Winzor DJ (1975) Further evidence for the concept of bovine plasma arylesterase as a lipoprotein. Biochem J 151: 625–630Google Scholar
  16. Doom JA, Sorenson RC, Billecke SS, Hsu C, La Du BN (1999) Evidence that several conserved histidine residues are required for hydrolytic activity of human paraoxonase/arylesterase. Chem. Biol. Interact 119-120: 235–241CrossRefGoogle Scholar
  17. Draganov DI, Stetson PL, Watson CE, Billecke SS, La Du BN (2000) Rabbit serum paraoxonase 3 (PON3) is an HDL-associated lactonase and protects LDL against oxidation. J Biol Chem 275: 33435–33442CrossRefGoogle Scholar
  18. Eckerson HW, Wyte CM, La Du BN (1983) The human serum paraoxonase/arylesterase polymorphism. Am J Hum Genet 35: 1126–1138Google Scholar
  19. Erdös EG, Debay CR, Westerman MP (1960) Arylesterases in blood:effect of calcium and inhibitors. Biochem Pharmacol 5: 173–186CrossRefGoogle Scholar
  20. Evans CH (1990) Chapter 2:Chemical properties of biochemical relevance In: Biochemistry of the lanthanides. Evans, CH (ed) Plenum Press, New York, NY, 9–46Google Scholar
  21. Furlong CE, Costa LG, Hassett C, Richter RJ, Sundstrom JA, Adler DA, Disteche CM, Omiecinski CJ, Chapline C, Crabb JW (1993) Human and rabbit paraoxonases: purification, cloning, sequencing, mapping and role of polymorphism in organophosphate detoxification. Chem Biol Interact 87: 35–48CrossRefGoogle Scholar
  22. Furlong CE, Richter RJ, Chapline C, Crabb JW (1991) Purification of rabbit and human serum paraoxonase. Biochemistry, 30: 10133–10140CrossRefGoogle Scholar
  23. Furlong CE, Richter RJ, Seidel SL, Motulsky AG (1988) Role of genetic polymorphism of human plasma paraoxonase/arylesterase in hydrolysis of the insecticide metabolites chlorpyrifos oxon and paraoxon. Am J Hum Genet 43: 230–238Google Scholar
  24. Gan KN, Smolen A, Eckerson HW, La Du BN (1991) Purification of human serum paraoxonase/arylesterase. Evidence for one esterase catalyzing both activities. Drug Metab Dispos 19: 100–106Google Scholar
  25. Garavito RM, Picot D, Loll PJ (1994) Prostaglandin H synthase. Curr Opin Struct Biol 4: 529–535CrossRefGoogle Scholar
  26. Garin MC, James RW, Dussoix P, Blanche H, Passa P, Froguel P, Ruiz J (1997) Paraoxonase polymorphism Met-Leu54 is associated with modified serum concentrations of the enzyme. A possible link between the paraoxonase gene and increased risk of cardiovascular disease in diabetes. J Clin Invest 99: 62–66CrossRefGoogle Scholar
  27. Geldmacher-Von Mallinckrodt M, Diepgen TL (1988) The human serum paraoxonase-Polymorphism and specificity. Toxicol Env Chem 18: 79–196CrossRefGoogle Scholar
  28. Goyal J, Wang K, Liu M, Subbaiah PV (1997) Novel Function of lecithin-cholesterol acyltransferase. J Biol Chem 272: 16231–16239CrossRefGoogle Scholar
  29. Hassett C, Richter RJ, Humbert R, Chapline C, Crabb JW, Omiecinski CJ, Furlong CE (1991) Characterization of cDNA clones encoding rabbit and human serum paraoxonase: the mature protein retains its signal sequence. Biochemistry 30: 10141–10149CrossRefGoogle Scholar
  30. Humbert R, Adler DA, Disteche CM, Hassett C, Omiecinski CJ, Furlong CE (1993) The molecular basis of the human serum paraoxonase activity polymorphism. Nat Genet 3: 73–76CrossRefGoogle Scholar
  31. Jakubowski H (2000) Calcium-dependent human serum homocysteine thiolactone hydrolase. A protective mechanism against protein N-homocysteinylation. J Biol Chem 275: 3957–3962CrossRefGoogle Scholar
  32. James RW, Blatter Garin MC, Calabresi L, Miccoli R, Von Eckardstein A, Tilly-Kiesi M, Taskinen MR, Assmann G, Franceschini G (1998) Modulated serum activities and concentrations of paraoxonase in high density lipoprotein deficiency states. Atherosclerosis 139: 77–82CrossRefGoogle Scholar
  33. Josse D (1999) Vers la détermination du centre actif et de la structure de la paraoxonase humaine. Ph D. dissertation. University de Grenoble, FranceGoogle Scholar
  34. Josse D, Xie W, Masson P, Schopfer M, Lockridge O (1999b) Tryptophan residue(s) as major components of the human serum paraoxonase active site. Chem Biol Interact, 119-120: 79–84CrossRefGoogle Scholar
  35. Josse D, Xie W, Renault F, Rochu D, Schopfer LM, Masson P, Lockridge O (1999a) Identification of residues essential for human paraoxonase (PON1) arylesterase/ organophosphatase activities. Biochemistry 38: 2816–2825CrossRefGoogle Scholar
  36. Kelso GJ, Stuart WD, Richter RJ, Furlong CE, Jordan-Starck TC, Harmony JA (1994) Apolipoprotein J is associated with paraoxonase in human plasma. Biochemistry 33: 832–839CrossRefGoogle Scholar
  37. Kitchen BJ, Masters CJ, Winzor DJ (1973) Effects of lipid removal on the molecular size and kinetic properties of bovine plasma arylesterase. Biochem J 135: 93–99Google Scholar
  38. Kobayashi M, Shinohara M, Sakoh C, Kataoka M, Shimizu S (1998) Lactone-ring-cleaving enzyme: genetic analysis, novel RNA editing, and evolutionary implications. Proc Natl Acad Sci USA 95: 12787–12792CrossRefGoogle Scholar
  39. Kuo CL, La Du BN (1998) Calcium binding by human and rabbit serum paraoxonases. Structural stability and enzymatic activity. Drug Metab Dispos, 26: 653–660Google Scholar
  40. Kuo CL, La Du BN (1995) Comparison of purified human and rabbit serum paraoxonases. Drug Metab Dispos 23: 935–944Google Scholar
  41. La Du BN, Aviram M, Billecke S, Navab M, Primo-Parmo S, Sorenson RC, Standiford TJ (1999) On the physiological role(s) of the paraoxonases. Chem Biol Interact, 119-120: 379–388CrossRefGoogle Scholar
  42. Mackness MI, Walker CH (1988) Multiple forms of sheep serum A-esterase activity associated with the high-density lipoprotein. Biochem J 250: 539–545Google Scholar
  43. Mackness MI, Walker CH (1983) Partial purification and properties of sheep serum“A’-esterases. Biochem Pharmacol 32: 2291–2296CrossRefGoogle Scholar
  44. Main AR (1960) The purification of the enzyme hydrolysing diethyl p-nitrophenyl phosphate (paraoxon) in sheep serum. Biochem J 74: 10–20Google Scholar
  45. Masson P, Josse D, Lockridge O, Viguté N, Taupin C, Buhler C (1998) Enzymes hydrolyzing organophosphates as potential catalytic scavengers against organophosphate poisoning. Physiol Paris 92: 357–362CrossRefGoogle Scholar
  46. Mazur A (1946) An enzyme in animal tissues capable of hydrolyzing the phosphorus-fluorine bond of alkyl fluorophosphates. J Biol Chem 164: 271–289Google Scholar
  47. McPhalen CA, Strynadka NCJ, James MNG (1991) Calcium-binding sites in proteins: a structural perspective. Adv Prot Chem 42: 77–144CrossRefGoogle Scholar
  48. Ozols J (1999) Isolation and complete covalent structure of liver microsomal paraoxonase. Biochem J, 338: 265–272CrossRefGoogle Scholar
  49. Primo-Parmo SL, Sorenson RC, Teiber J, La Du BN (1996) The human serum paraoxonase/ arylesterase gene (PON1) is one member of a multigene family. Genomics 33: 498–507CrossRefGoogle Scholar
  50. Segrest JP, Darber DW, Brouillette CG, Harvey SC, Anantharamaiah GM (1994) The amphipathic a-helix: amultifunctional structural motif in plasma apolipoproteins. Adv Prot Chem 45: 303–369CrossRefGoogle Scholar
  51. Smolen A, Eckerson HW, Gan KN, Hailat N, La Du BN (1991) Characteristics of the genetically determined allozymic forms of human serum paraoxonase/arylesterase. Drug Metab Dispos 19: 107–112Google Scholar
  52. Sorenson RC, Aviram M, Bisgaier CL, Billecke S, Hsu C, La Du BN (1999) Properties of the retained N-terminal hydrophobic leader sequence in human serum paraoxonase/ arylesterase. Chem Biol Interact, 119-120: 243–249CrossRefGoogle Scholar
  53. Sorenson RC, Primo-Parmo SL, Kuo CL, Adkins S, Lockridge O, La Du BN (1995) Reconsideration of the catalytic center and mechanism of mammalian paraoxonase/ arylesterase. Proc. Natl Acad Sci USA 92: 7187–7191CrossRefGoogle Scholar
  54. Swaney JB, O’Brien K (1978) Cross-linking studies of the self-association properties of apo A-I and apo A-II from human high density lipoprotein. J Biol Chem 253: 7069–7077Google Scholar
  55. Vallee BL, Auld DS (1993) New perspective on zinc biochemistry: cocatalytic sites in multizinc enzymes. Biochemistry 32: 6493–6500CrossRefGoogle Scholar
  56. Vohl M.-C, Neuville TA-M, Kumarathasan R, Braschi S, Sparks DL (1999) A novel lecithin-cholesterol acyltransferase antioxidant activity prevents the formation of oxidized lipids during lipoprotein oxidation. Biochemistry 38: 5976–5981CrossRefGoogle Scholar
  57. Watson AD, Berliner JA, Hama SY, La Du BN, Faull KF, Fogelman AM, Navab M (1995) Protective effect of high density lipoprotein associated paraoxonase. Inhibition of the biological activity of minimally oxidized low density lipoprotein. J Clin Invest 96: 2882–2891CrossRefGoogle Scholar
  58. Wendt KU, Lenhart A, Schulz GE (1999) The structure of the membrane protein squalenehopene cyclase at 2.0 Å resolution. J. Mol. Biol 286: 175–187CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2002

Authors and Affiliations

  • Denis Josse
    • 1
  • Patrick Masson
    • 1
  • Cynthia Bartels
    • 2
  • Oksana Lockridge
    • 2
  1. 1.Unité d’enzymologieCentre de Recherches du Service de Santé des ArméesLa Tronche CedexFrance
  2. 2.Eppley InstituteUniversity of Nebraska Medical CenterOmahaUSA

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