The reconstituted ADP/ATP carrier can mediate H+ transport by free fatty acids, which is further stimulated by mersalyl

Nikolai Broustovetski, Martin Klingenberg

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

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 Δψ in AAC-cytochrome-c oxidase (COX)-coreconstituted proteoliposomes. FFA increased the initial rate of proton transport in AAC proteoliposomes and decreased Δψ 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 Δψ 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 unidirectional transport mode.

Original languageEnglish (US)
Pages (from-to)27329-27336
Number of pages8
JournalJournal of Biological Chemistry
Volume269
Issue number44
StatePublished - Nov 4 1994
Externally publishedYes

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Mersalyl
ATP Translocases Mitochondrial ADP
Nonesterified Fatty Acids
Protons
Oxidoreductases
Membranes
Guanine Nucleotides
Electron Transport Complex IV
Membrane Potentials

ASJC Scopus subject areas

  • Biochemistry

Cite this

The reconstituted ADP/ATP carrier can mediate H+ transport by free fatty acids, which is further stimulated by mersalyl. / Broustovetski, Nikolai; Klingenberg, Martin.

In: Journal of Biological Chemistry, Vol. 269, No. 44, 04.11.1994, p. 27329-27336.

Research output: Contribution to journalArticle

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