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(Received for publication, July 23, 1996, and in revised form, August 20, 1996)
From the Department of Health Chemistry, School of Pharmaceutical
Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa-ku,
Tokyo 142, Japan
Type II secretory phospholipase A2
(sPLA2) has been shown to be induced by a variety of
proinflammatory stimuli and, therefore, has been implicated in the
inflammatory process. In order to determine whether association of
sPLA2 with cell surfaces via heparan sulfate proteoglycan
is important for its effects on cellular functions, we have identified
the critical domain in sPLA2 for heparin and cell surface
binding and examined its role in cellular prostaglandin (PG)
biosynthesis. Replacement of several conserved Lys residues in the
C-terminal region of mouse and rat sPLA2s by Glu resulted in a marked reduction of their capacities to bind to heparin and mammalian cell surfaces without affecting their enzymatic activities toward dispersed phospholipid as a substrate. CHO cells stably transfected with wild-type sPLA2 released about twice as
much arachidonic acid (AA) during culture for 10 h with fetal calf serum and interleukin-1
Volume 271, Number 47,
Issue of November 22, 1996
pp. 30041-30051
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
than cells transfected with vector alone, whereas the ability to enhance AA release was impaired in
sPLA2 mutants incapable of binding to cell surfaces. AA
released by wild-type sPLA2-transfected CHO cells was
metabolized to prostaglandin E2 via prostaglandin
endoperoxide H synthase (PGHS)-2 after IL-1
stimulation, revealing a
particular functional linkage of sPLA2 to PGHS-2. In
contrast, A23187-initiated immediate AA release over 30 min was not
affected by sPLA2 overexpression. Taken together, these
results suggest that sPLA2 expressed endogenously and
anchored on cell surfaces via its C-terminal heparin-binding domain is involved in the PGHS-2-dependent delayed PG biosynthesis
initiated by growth factors and cytokines during long term culture.
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