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J Biol Chem, Vol. 274, Issue 34, 23940-23947, August 20, 1999
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From the We examined the actions of sphingosine
1-phosphate (S1P) on signaling pathways in Chinese hamster ovary cells
transfected with putative S1P receptor subtypes, i.e.
Edg-1, AGR16/H218 (Edg-5), and Edg-3. Among these receptor-transfected
cells, there was no significant difference in the expressing numbers of
the S1P receptors and their affinities to S1P, which were estimated by
[3H]S1P binding to the cells. In vector-transfected
cells, S1P slightly increased cytosolic Ca2+ concentration
([Ca2+]i) in association with inositol phosphate
production, reflecting phospholipase C activation; the S1P-induced
actions were markedly enhanced in the Edg-3-transfected cells and
moderately so in the AGR16-transfected cells. In comparison with
vector-transfected cells, the S1P-induced [Ca2+]i
increase was also slightly enhanced in the Edg-1-transfected cells. In
all cases, the inositol phosphate and Ca2+ responses to S1P
were partially inhibited by pertussis toxin (PTX). S1P also
significantly increased cAMP content in a PTX-insensitive manner in all
the transfected cells; the rank order of their intrinsic activity of
S1P receptor subtypes was AGR16 > Edg-3 > Edg-1. In the
presence of forskolin, however, S1P significantly inhibited cAMP
accumulation at a lower concentration (1-100 nM) of S1P in a manner sensitive to PTX in the Edg-1-transfected cells but not in
either the Edg-3 or AGR16-transfected cells. As for cell migration activity evaluated by cell number across the filter of blind Boyden chamber, Edg-1 and Edg-3 were equally potent, but AGR16 was
ineffective. Thus, S1P receptors may couple to both PTX-sensitive and
-insensitive G-proteins, resulting in the selective regulation of the
phospholipase C-Ca2+ system, adenylyl cyclase-cAMP system,
and cell migration activity, according to the receptor subtype.
Laboratory of Signal Transduction,
Tokyo Metropolitan Institute of Medical
Science,
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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