Exploration of the nucleotide binding sites of the isolated ADP/ATP carrier protein from beef heart mitochondria. 2. Probing of the nucleotide sites by formycin triphosphate, a fluorescent transportable analog of ATP

Brandolin, Gérard; Dupont, Yves; Vignais, Pierre V.

doi:10.1021/bi00268a005

Cited by 19 publications

(8 citation statements)

References 7 publications

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“…This intermolecular mechanism implies the existence of positive interactions between distinct binding sites exposed to the outer and inner face of the membrane-embedded carrier. Consistent with these findings, the occurrence of distinct specific nucleotide-binding sites on the ADP/ATP carrier was deduced from the binding of ADP and ATP derivatives to the carrier either in the membrane-bound state or when isolated in detergent solution (60)(61)(62). For example, in the mitochondrial membrane, two specific nucleotide-binding sites are located on the same face of the membrane, either the IMS face of intact mitochondria or the matrix face of inside-out particles.…”

Section: What Triggers the Changes? Kinetic Aspectsmentioning

confidence: 56%

Relations Between Structure and Function of the Mitochondrial ADP/ATP Carrier

Nury

Dahout-Gonzalez

Trezeguet

et al. 2006

Annu. Rev. Biochem.

149

139

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Import and export of metabolites through mitochondrial membranes are vital processes that are highly controlled and regulated at the level of the inner membrane. Proteins of the mitochondrial carrier family ( MCF ) are embedded in this membrane, and each member of the family achieves the selective transport of a specific metabolite. Among these, the ADP/ATP carrier transports ADP into the mitochondrial matrix and exports ATP toward the cytosol after its synthesis. Because of its natural abundance, the ADP/ATP carrier is the best characterized within MCF, and a high-resolution structure of one conformation is known. The overall structure is basket shaped and formed by six transmembrane helices that are not only tilted with respect to the membrane, but three of them are also kinked at the level of prolines. The functional mechanisms, nucleotide recognition, and conformational changes for the transport, suggested from the structure, are discussed along with the large body of biochemical and functional results.

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Section: What Triggers the Changes? Kinetic Aspectsmentioning

confidence: 56%

Relations Between Structure and Function of the Mitochondrial ADP/ATP Carrier

Nury

Dahout-Gonzalez

Trezeguet

et al. 2006

Annu. Rev. Biochem.

149

139

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“…It is probable that the effect of ATP and ADP on the intrinsic fluorescence of the carrier is shared by other transportable nucleotides. In fact, it was previously reported that formycin triphosphate, a transportable ATP analogue that binds to the isolated carrier is displaced by BA (Brandolin et al, 1982) in contrast to N-ATP, an analogue that binds to the carrier but is not transported (Dupont et al, 1982). On the basis of modifications of its fluorescence properties upon addition of CATR in the absence of nucleotides, the isolated carrier was found to bind CATR.…”

Section: Discussionmentioning

confidence: 97%

“…Although appealing, the tetrameric hypothesis is apparently in conflict with structural studies showing that the CATRand BA-carrier complexes behave as dimers (Kramer & Klingenberg, 1977;Block et al, 1982a). However, recent binding studies on the isolated AdN carrier (Dupont et al, 1982;Brandolin et al, 1982) and on the membrane-bound carrier (Block & Vignais, 1984) tend to support the tetramer hypothesis. The tetrameric organization of the AdN carrier is also consistent with the combined observations that there is more than one CATR binding site per transport unit (Figure 7) and that CATR binds to one subunit only of the carrier dimer (Kramer & Klingenberg, 1977;Block et al, 1981).…”

Section: Discussionmentioning

confidence: 99%

“…However, an alternative explanation is that discrete molecular events occur in the course of the transition from the CATR conformation to the BA conformation. In this context, it may be recalled that by means of titration with fluorescent nucleotides, four nucleotide binding sites have been demonstrated in the isolated carrier (Dupont et al, 1982;Brandolin et al, 1982). It is plausible that the four nucleotide sites are distributed in distinct subunits of a tetramer.…”

Section: Discussionmentioning

confidence: 99%

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Substrate-induced modifications of the intrinsic fluorescence of the isolated adenine nucleotide carrier protein: demonstration of distinct conformational states

Brandolin¹,

Dupont²,

Vignais³

1985

Biochemistry

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The effects of ATP or ADP and the specific inhibitors carboxyatractyloside (CATR) and bongkrekic acid (BA) on the conformation of the isolated adenine nucleotide (AdN) carrier protein were studied by fluorescence spectroscopy. The addition of ATP to the AdN carrier resulted in a rapid fluorescence increase of the tryptophanyl residue(s) at 355 nm, which leveled up in less than 1 s at 22 degrees C. Among the natural nucleotides, only ATP and ADP were effective. At 10 degrees C or below, the kinetics of the fluorescence increase induced by ATP were biphasic, consisting of a rapid phase of less than 1 s, followed by a slower phase that lasted for a few seconds and had virtually the same amplitude as the rapid one. Both phases were abolished when CATR was added prior to ATP or fully reversed when CATR was added after the fluorescence response to ATP had been elicited. The number of CATR binding sites present on the carrier protein was determined by CATR specific inhibition of the ATP-induced increase in intrinsic fluorescence. The calculated number of CATR sites was equal to that found by another method based on the use of the same preparation of AdN carrier loaded with fluorescent nucleotide naphthoyl-ATP and on the CATR-induced release of the bound naphthoyl-ATP, demonstrating the reliability of the intrinsic fluorescence assay. Addition of BA prior to or together with ATP nearly doubled the amplitude of the ATP-induced fluorescence signal. At 10 degrees C or below, the fluorescence response to ATP in the presence of BA could also be decomposed into rapid and slow phases.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…In addition, experimental evidence for a dimeric organization of other MCF members, such as the oxaloglutarate, citrate, or Pi carriers, support the hypothesis of an oligomerization of the mitochondrial carriers [64,65]. A tetrameric functional unit of Aacp was also suggested because two nucleotide binding sites on each side of the carrier and of different affinities were characterized for one transport unit [66,67]. However, recent findings questioned the existence of an oligomeric state of MCF members.…”

Section: The Different Proteins Involved In the Mitochondrial Membmentioning

confidence: 99%

Yeast Mitochondrial Interactosome Model: Metabolon Membrane Proteins Complex Involved in the Channeling of ADP/ATP

Clémençon

2012

IJMS

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The existence of a mitochondrial interactosome (MI) has been currently well established in mammalian cells but the exact composition of this super-complex is not precisely known, and its organization seems to be different from that in yeast. One major difference is the absence of mitochondrial creatine kinase (MtCK) in yeast, unlike that described in the organization model of MI, especially in cardiac, skeletal muscle and brain cells. The aim of this review is to provide a detailed description of different partner proteins involved in the synergistic ADP/ATP transport across the mitochondrial membranes in the yeast Saccharomyces cerevisiae and to propose a new mitochondrial interactosome model. The ADP/ATP (Aacp) and inorganic phosphate (PiC) carriers as well as the VDAC (or mitochondrial porin) catalyze the import and export of ADP, ATP and Pi across the mitochondrial membranes. Aacp and PiC, which appear to be associated with the ATP synthase, consist of two nanomotors (F0, F1) under specific conditions and form ATP synthasome. Identification and characterization of such a complex were described for the first time by Pedersen and co-workers in 2003.

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Exploration of the nucleotide binding sites of the isolated ADP/ATP carrier protein from beef heart mitochondria. 2. Probing of the nucleotide sites by formycin triphosphate, a fluorescent transportable analog of ATP

Cited by 19 publications

References 7 publications

Relations Between Structure and Function of the Mitochondrial ADP/ATP Carrier

Relations Between Structure and Function of the Mitochondrial ADP/ATP Carrier

Substrate-induced modifications of the intrinsic fluorescence of the isolated adenine nucleotide carrier protein: demonstration of distinct conformational states

Yeast Mitochondrial Interactosome Model: Metabolon Membrane Proteins Complex Involved in the Channeling of ADP/ATP

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