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J. Biol. Chem., Vol. 275, Issue 44, 34534-34540, November 3, 2000
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From the Department of Medicine and Canadian Institutes for Health
Research Group on Molecular and Cell Biology of Lipids, University of
Alberta, Edmonton, Alberta T6G 2S2, Canada
We report the subcellular localization of enzymes
involved in phosphatidylserine biosynthesis in mammalian cells. Several lines of evidence suggest that phosphatidylserine synthase-1 (PSS1) is
highly enriched in mitochondria-associated membranes (MAM) and is
largely excluded from the bulk of the endoplasmic reticulum (ER).
Taking advantage of the substrate specificity of PSS1, we showed that
(i) MAM contain choline exchange activity, whereas this activity is
very low in the bulk of the ER, (ii) serine exchange activity is
inhibited by choline to a much greater extent in MAM than in ER, and
(iii) MAM use phosphatidylcholine and phosphatidylethanolamine as
substrates for phosphatidylserine biosynthesis, whereas the ER utilizes
only phosphatidylethanolamine. According to immunoblotting of proteins
from both CHO-K1 cells and murine liver, PSS1 is localized to MAM, and
in hepatoma cells stably expressing PSS1 this protein is highly
enriched in MAM. Since the ER contains serine and ethanolamine exchange
activities, we had predicted that PSS2 would account for the serine
exchange activity in the ER. Unexpectedly, using immunoblotting
experiments, we found that (i) PSS2 of CHO-K1 cells is present only in
MAM and (ii) PSS2 is restricted to MAM of McArdle cells expressing
recombinant PSS2. These data leave open the question of which enzyme
imparts PSS activity to the ER and suggest that a third isoform of PSS
might be located in the ER.
Phosphatidylserine Synthase-1 and -2 Are Localized to
Mitochondria-associated Membranes*
*
This work was supported by an operating grant (to
J. E. V.) and a studentship (to S. J. S.) from the Medical Research
Council of Canada.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: 332 Heritage Medical
Research Center, University of Alberta, Edmonton, Alberta T6G 2S2,
Canada. Tel.: 780-492-7250; Fax: 780-492-3383; E-mail: jean.vance@ualberta.ca.
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