Kinetics of electrogenic transport by the ADP/ATP carrier

T. Gropp, Nikolai Broustovetski, M. Klingenberg, V. Müller, K. Fendler, Ernst Bamberg

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

The electrogenic transport of ATP and ADP by the mitochondrial ADP/ATP carrier (AAC) was investigated by recording transient currents with two different techniques for performing concentration jump experiments: 1) the fast fluid injection method: AAC-containing proteoliposomes were adsorbed to a solid supported membrane (SSM), and the carrier was activated via ATP or ADP concentration jumps. 2) BLM (black lipid membrane) technique: proteoliposomes were adsorbed to a planar lipid bilayer, while the carrier was activated via the photolysis of caged ATP or caged ADP with a UV laser pulse. Two transport modes of the AAC were investigated, ATP(ex)-0(in) and ADP(ex)-0(in). Liposomes not loaded with nucleotides allowed half-cycles of the ADP/ATP exchange to be studied. Under these conditions the AAC transports ADP and ATP electrogenically. Mg2+ inhibits the nucleotide transport, and the specific inhibitors carboxyatractylate (CAT) and bongkrekate (BKA) prevent the binding of the substrate. The evaluation of the transient currents yielded rate constants of 160 s-1 for ATP and ≥400 s-1 for ADP translocation. The function of the carrier is approximately symmetrical, i.e., the kinetic properties are similar in the inside-out and right-side-out orientations. The assumption from previous investigations, that the deprotonated nucleotides are exclusively transported by the AAC, is supported by further experimental evidence. In addition, caged ATP and caged ADP bind to the carrier with similar affinities as the free nucleotides. An inhibitory effect of anions (200-300 mM) was observed, which can be explained as a competitive effect at the binding site. The results are summarized in a transport model.

Original languageEnglish (US)
Pages (from-to)714-726
Number of pages13
JournalBiophysical Journal
Volume77
Issue number2
StatePublished - Aug 1999
Externally publishedYes

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ATP Translocases Mitochondrial ADP
Adenosine Diphosphate
Adenosine Triphosphate
Nucleotides
Bongkrekic Acid
Photolysis
Lipid Bilayers
Membrane Lipids
Liposomes
Anions
Lasers
Binding Sites
Injections
Membranes

ASJC Scopus subject areas

  • Biophysics

Cite this

Gropp, T., Broustovetski, N., Klingenberg, M., Müller, V., Fendler, K., & Bamberg, E. (1999). Kinetics of electrogenic transport by the ADP/ATP carrier. Biophysical Journal, 77(2), 714-726.

Kinetics of electrogenic transport by the ADP/ATP carrier. / Gropp, T.; Broustovetski, Nikolai; Klingenberg, M.; Müller, V.; Fendler, K.; Bamberg, Ernst.

In: Biophysical Journal, Vol. 77, No. 2, 08.1999, p. 714-726.

Research output: Contribution to journalArticle

Gropp, T, Broustovetski, N, Klingenberg, M, Müller, V, Fendler, K & Bamberg, E 1999, 'Kinetics of electrogenic transport by the ADP/ATP carrier', Biophysical Journal, vol. 77, no. 2, pp. 714-726.
Gropp T, Broustovetski N, Klingenberg M, Müller V, Fendler K, Bamberg E. Kinetics of electrogenic transport by the ADP/ATP carrier. Biophysical Journal. 1999 Aug;77(2):714-726.
Gropp, T. ; Broustovetski, Nikolai ; Klingenberg, M. ; Müller, V. ; Fendler, K. ; Bamberg, Ernst. / Kinetics of electrogenic transport by the ADP/ATP carrier. In: Biophysical Journal. 1999 ; Vol. 77, No. 2. pp. 714-726.
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