Abstract
Serum paraoxonase (PON) is associated with plasma high density lipoproteins, and prevents the oxidative modification of low density lipoproteins. We have developed a sensitive sandwich enzyme-linked immunosorbent assay (ELISA), using two monoclonal antibodies against PON, to measure serum PON concentration. The concentration of PON in healthy Japanese subjects was 59.3 +/- 1.3 microgram/mL (mean +/- SEM; n = 87). Serum PON concentrations in relation to the PON 192 genetic polymorphism were: 69.5 +/- 2.9 microgram/mL in the QQ genotype; 63.0 +/- 1.9 microgram/mL in the QR genotype; and 52.8 +/- 1.7 microgram/mL in the RR genotype. Concentrations were significantly lower in the RR than in the QQ genotype (P < 0.01). Serum paraoxonase specific activity was higher in RR than in QQ subjects (18.6 +/- 0.40 vs. 2. 56 +/- 0.05 nmol/min/microgram, P < 0.01), but arylesterase specific activity was unrelated to genotype. PON concentration was positively associated (P < 0.001) with both serum arylesterase activity and, after adjusting for the effect of the position 192 polymorphism, with serum paraoxonase activity. Subjects with angiographically verified coronary heart disease had significantly lower PON concentrations than the healthy controls (52.0 +/- 2.3 microgram/mL; n = 35, P < 0.01). This association was independent of the position 192 genotype. Our new ELISA should be of value for epidemiologic and clinical studies of serum PON concentration. immunosorbent assay for human serum paraoxonase concentration.
Highlights
Serum paraoxonase (PON) is associated with plasma high density lipoproteins, and prevents the oxidative modification of low density lipoproteins
Its natural substrate is not certain, studies have indicated that PON hydrolyzes phospholipid peroxides in both high density lipoproteins (HDLs) and low density lipoproteins (LDLs) [2], and that this is one mechanism by which HDLs prevent the oxidative modification of LDLs [3]
Low serum paraoxonase activity has been observed in patients with myocardial infarction [8], hypercholesterolemia [9], and diabetes mellitus [9], and there is evidence that the position 192 polymorphism may be a risk factor for coronary heart disease (CHD) in some populations [10]
Summary
Serum paraoxonase (PON) is associated with plasma high density lipoproteins, and prevents the oxidative modification of low density lipoproteins. We have developed a sensitive sandwich enzyme-linked immunosorbent assay (ELISA), using two monoclonal antibodies against PON, to measure serum PON concentration. Subjects with angiographically verified coronary heart disease had significantly lower PON concentrations than the healthy controls (52.0 ؎ 2.3 g/mL; n ؍35, P Ͻ 0.01). This association was independent of the position 192 genotype. A sandwich enzyme-linked immunosorbent assay for human serum paraoxonase concentration. Propanesulfonic acid; CHD, coronary heart disease; EDTA, ethylenediamine-N,N,NЈ,NЈ-tetraacetic acid; ELISA, enzyme-linked immunosorbent assay; LCAT, lecithin–cholesterol acyltransferase; PAF-AH, plateletactivating factor acetylhydrolase; PON, paraoxonase.
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