Spectrophotometric Assay for Serum Platelet-activating Factor Acetylhydrolase Activity

Overview

Journal Clin Chim Acta

Specialty Biochemistry

Date 2000 May 16

PMID 10807978

Citations 17

Authors

T Kosaka

M Yamaguchi

Y Soda

T Kishimoto

A Tago

M Toyosato

K Mizuno

Affiliations

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Abstract

We developed a spectrophotometric assay for serum platelet-activating factor acetylhydrolase (PAF-AH, EC 3.1.1.47.) activity using a platelet-activating factor (PAF) analogue with a 4-nitrophenyl group as substrate. PAF-AH hydrolyzes the sn-2 position of the substrate ¿1-myristoyl-2-(p-nitrophenylsuccinyl)phosphatidylcholine, producing p-nitrophenyl succinate. This liberation was spectrophotometrically monitored and the activity determined from the change in absorption. The assay does not require radioisotopes and is applicable to an automatic analyzer. Utilizing this assay with an automatic analyzer, it is possible to measure the activities of thousands of samples in a few hours with excellent precision (CV 0.5%, n=30) and high correlation (r=0.979, n=100) with the results of a conventional radioisotopic assay. The assay should be particularly useful for clinical diagnostics.

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