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J Biol Chem, Vol. 273, Issue 41, 26638-26644, October 9, 1998
From the Department of Physiology, Tufts University School of
Medicine, Boston, Massachusetts 02111 and the ¶ Department of
Biomedical Research, St. Elizabeth's Medical Center, Boston,
Massachusetts 02135
Recent studies indicate that wortmannin, a potent
inhibitor of phosphatidylinositol (PI) 3-kinase, interferes with bile
acid secretion in rat liver; taurocholate induces recruitment of
ATP-dependent transporters to the bile canalicular
membrane, and PI 3-kinase products are important in intracellular
trafficking.
We investigated the role of PI 3-kinase in bile acid secretion by
studying the in vivo effect of taurocholate, colchicine, and wortmannin on bile acid secretion, kinase activity, and protein levels in canalicular membrane vesicle (CMV) and sinusoidal membrane vesicle (SMV) fractions from rat liver. Treatment of rats or perfusion of isolated liver with taurocholate significantly increased PI 3-kinase
activity in both membrane fractions. Taurocholate increased protein
content of ATP-dependent transporters, which were detected only in CMVs, whereas increased levels of p85 and a cell adhesion molecule, cCAM 105, were observed in both fractions.
Colchicine prevented taurocholate-induced changes in all proteins
studied, as well as the increase in PI 3-kinase activity in CMVs, but
it resulted in further accumulation of PI 3-kinase activity, p85, and
cCAM 105 in SMVs. These results indicate that taurocholate-mediated
changes involve a microtubular system.
Wortmannin blocked taurocholate-induced bile acid secretion. The effect
was more profound when wortmannin was administered prior to treatment
with taurocholate. When wortmannin was given after taurocholate, the
protein levels of each ATP-dependent transporter were
maintained in CMVs, whereas the levels of p85 and cCAM decreased in
both membrane fractions. Perfusion of liver with wortmannin before
taurocholate administration blocked accumulation of all proteins
studied in CMVs and SMVs.
These results indicate that PI 3-kinase is required for intracellular
trafficking of itself, as well as of ATP-dependent
canalicular transporters.
The Role of Phosphoinositide 3-Kinase in Taurocholate-induced
Trafficking of ATP-dependent Canalicular Transporters in
Rat Liver
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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