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J. Biol. Chem., Vol. 269, Issue 44, 27329-27336, Nov, 1994
N Brustovetsky and M Klingenberg
In a reconstituted system, the participation of the ATP/ADP carrier (AAC)
in the free fatty acid (FFA)-induced proton transport was demonstrated (i)
by direct measuring of the proton transport through the membranes of AAC
proteoliposomes and (ii) by monitoring of the transmembrane potential delta
psi in AAC-cytochrome-c oxidase (COX)- coreconstituted proteoliposomes. FFA
increased the initial rate of proton transport in AAC proteoliposomes and
decreased delta psi in AAC- COX proteoliposomes. Inhibitors of AAC
suppressed the effects of FFA. Without AAC or with inactive AAC, FFA cannot
maintain proton leakage through the membrane. In these cases, even a small
increase of delta psi was induced by FFA. These results demonstrate for the
first time with purified components a participation of AAC in FFA-induced
proton transport supporting an earlier suggestion (Skulachev, V.P. (1991)
FEBS Lett. 294, 158-162). Mersalyl treatment of the AAC-COX proteoliposomes
resulted in an increase of the AAC-mediated protonophoric action of FFA.
Mersalyl also sensitized the protonophoric action of the FFA against
nucleotides so that even guanine nucleotides, which are inactive in
transport, become inhibitory. The effect of mersalyl is rationalized in
terms of a specific interaction with cysteine 159 being attracted as anion
by surrounding positive charges. This might open a gate similarly as
suggested for eosin 5-maleimide interaction (Majima, E., Koike, H., Hong,
Y.-M., Shinohara, Y., and Terada, H. (1993) J. Biol. Chem. 268,
22181-22187) and, thus, transform the AAC into undirectional transport
mode.
The reconstituted ADP/ATP carrier can mediate H+ transport by free fatty acids, which is further stimulated by mersalyl
Institute of Physical Biochemistry, University of Munich, Federal Republic of Germany.
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