Comparison of purified human and rabbit serum paraoxonases.
Rabbit serum paraoxonase (PON) activity was reported to be nearly 20 times greater than that found for humans and all other mammalian species tested, to date. However, 85% of the amino acid residues are identical in human and rabbit PONs, and the two purified PONs show similar substrate specificity patterns. Both are stimulated by phospholipids and have two asparagine-linked sugar chains. Both also have one intramolecular disulfide bond and one free sulfhydryl residue per molecule. Both require Ca2+ for stability and for catalytic activity. Zn2+ and Cd2+ also stabilize both PONs and prevent irreversible denaturation, but neither metal confers catalytic activity. Maximum specific activities for both esterases were approximately 2,000 units of arylesterase activity/mg protein. In contrast, rabbit PON is more stable than human PON to heat inactivation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis treatment. Chelex 100 strips Ca2+ from human PON more easily, and EDTA is less inhibitory with rabbit PON. We conclude that human and rabbit PONs have very similar active centers, but the latter binds Ca2+ more tightly, is a more stable enzyme, and is maintained at 3- to 4-fold higher steady-state concentrations in serum than its human counterpart. PON activity depends on adequate Ca2+ being available; therefore, apparently much higher levels of PON activity in rabbits can be explained by the reduced Ca2+ concentrations present in the early assay methods.
Kuo
C L
CL
Department of Environmental and Industrial Health, School of Public Health, University of Michigan Medical School, Ann Arbor 48109-0572, USA.
La Du
B N
BN
D000595:Amino Acid Sequence; D000818:Animals; D043303:Aryldialkylphosphatase; D002614:Chelating Agents / Q000494:pharmacology; D004220:Disulfides / Q000737:chemistry; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004950:Esterases / Q000037:antagonists & inhibitors / Q000097:blood* / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000494:pharmacology; D011817:Rabbits; D012117:Resins, Synthetic; D013045:Species Specificity; D013268:Stimulation, Chemical; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000737:chemistry
D002614:Chelating Agents; D004220:Disulfides; D010743:Phospholipids; D012117:Resins, Synthetic; D013438:Sulfhydryl Compounds; C024997:Chelex 100; D004492:Edetic Acid; D004950:Esterases; D043303:Aryldialkylphosphatase; D005947:Glucose