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Blood, 1 March 2002, Vol. 99, No. 5, pp. 1802-1810
PHAGOCYTES
Defects in leukocyte-mediated initiation of lipid peroxidation in
plasma as studied in myeloperoxidase-deficient subjects: systematic
identification of multiple endogenous diffusible substrates for
myeloperoxidase in plasma
Renliang Zhang,
Zhongzhou Shen,
William M. Nauseef, and
Stanley L. Hazen
From the Department of Cell Biology, the Department of
Cardiology, and the Center for Cardiovascular Diagnostics, Preventive
Cardiology Section, Cleveland Clinic Foundation, OH; the Chemistry
Department, Cleveland State University, OH; and the Inflammation
Program and Department of Medicine, University of Iowa and Veterans
Administration Medical Center, Iowa City.
More than a decade ago it was demonstrated that neutrophil
activation in plasma results in the time-dependent formation of lipid
hydroperoxides through an unknown, ascorbate-sensitive pathway. It is
now shown that the mechanism involves myeloperoxidase
(MPO)-dependent use of multiple low-molecular-weight substrates
in plasma, generating diffusible oxidant species. Addition of activated
human neutrophils (from healthy subjects) to plasma (50%, vol/vol)
resulted in the peroxidation of endogenous plasma lipids by catalase-,
heme poison-, and ascorbate-sensitive pathways, as assessed
by high-performance liquid chromatography (HPLC) with on-line
electrospray ionization tandem mass spectrometric analysis of
free and lipid-bound 9-HETE and 9-HODE. In marked contrast, neutrophils
isolated from multiple subjects with MPO deficiency failed to initiate
peroxidation of plasma lipids, but they did so after
supplementation with isolated human MPO. MPO-dependent use of a
low-molecular-weight substrate(s) in plasma for initiating lipid
peroxidation was illustrated by demonstrating that the filtrate of
plasma (10-kd MWt cutoff) could supply components required for
low-density lipoprotein lipid peroxidation in the presence of MPO and
H2O2. Subsequent HPLC fractionation of plasma
filtrate (10-kd MWt cutoff) by sequential column chromatography identified nitrite, tyrosine, and thiocyanate as major endogenous substrates and 17 -estradiol as a novel minor endogenous substrate in
plasma for MPO in promoting peroxidation of plasma lipids. These
results strongly suggest that the MPO-H2O2
system of human leukocytes serves as a physiological mechanism for
initiating lipid peroxidation in vivo.
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