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J. Biol. Chem., Vol. 276, Issue 15, 11461-11464, April 13, 2001
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,¶
From the Departments of Escherichia coli phospholipids
and lipopolysaccharide, made on the inner surface of the inner
membrane, are rapidly transported to the outer membrane by mechanisms
that are not well characterized. We now report a temperature-sensitive
mutant (WD2) with an A270T substitution in a trans-membrane region of
the ABC transporter MsbA. As shown by
32Pi and 14C-acetate labeling,
export of all major lipids to the outer membrane is inhibited by
~90% in WD2 after 30 min at 44 °C. Transport of newly synthesized
proteins is not impaired. Electron microscopy shows reduplicated inner
membranes in WD2 at 44 °C, consistent with a key role for MsbA in
lipid trafficking.
Biochemistry and
§ Cell Biology, Duke University Medical Center, Durham,
North Carolina 27710
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