Experimental Demonstration of the Reorienting (Mobile) Carrier Mechanism Exemplified by the Mitochondrial Adenine Nucleotide Translocator

Klingenberg, Martin; Scherer, B.; Stengel-Rutkowski, L.; Buchholz, Marlies; Grebe, K.

doi:10.1016/b978-0-12-068960-6.50024-1

Cited by 27 publications

(14 citation statements)

References 22 publications

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“…4) despite the nonlinear relations inherent in equation (1). In fact the slopes of these lines fairly well represent the Kd's for CAT and ATR: the ratio of the slopes is 3 1, is approximate agreement with the ratios of KiTR/eAT [4]. The results demonstrate that ATR' and CAT or ATR compete obviously for the same binding site at the carrier.…”

Section: North-holland Publishing Company -Amsterdamsupporting

confidence: 70%

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The new atractyloside type compound as a high affinity ligand to the adenine nucleotide carrier

et al. 1973

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Section: North-holland Publishing Company -Amsterdamsupporting

confidence: 70%

“…Furthermore BA was unable to remove CAT, obviously due to the higher affinity of CAT as compared to ATR. Thus BA furnished a very important means of discriminating between [35S]ATR and [35S]CAT [4]. The application of this method is shown in the experiment in fig.…”

Section: The Injluence Of Bongkrekic Acid On the Binding Of Atr'mentioning

confidence: 96%

The new atractyloside type compound as a high affinity ligand to the adenine nucleotide carrier

et al. 1973

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“…Palmityl-CoA had been shown to be an inhibitor of the ADP/ATP exchange [12,12a]. The effect of palmityl-CoA on mitochondria resembles atractylate more than bongkrekate with resepect to the influence on ADP binding and the contractibility of mitochondria [13]. As shown in Fig.…”

Section: The Efiect Of Inhibitors On the Adpiatp Exchangementioning

confidence: 78%

“…It is probable that only particles with inverted membranes should have trapped added ADP and ATP since the inversion process encloses external fluid, and thus be capable of exchange. It is not known what portion is inverted, a portion as high as 40% of right-sided particles has been estimated in context with the binding of atractylate to sonic particles [13].…”

Section: Discussionmentioning

confidence: 99%

The Adenine-Nucleotide Exchange in Submitochondrial (Sonic) Particles

Klingenberg

1977

Eur J Biochem

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Transport of adenine nucleotides through the inner mitochondrial membrane is investigated with submitochondrial (sonic) particles from beef heart mitochondria. Significant transport of nucleotides is observed only when the particles are loaded with adenine nucleotides during sonication. The nucleotide transport is characterized as follows.1. Sonic particles can be loaded during sonication in the presence of 20 mM ADP or ATP with up to 18 pmol/g protein, of a mixture of ATP, ADP and AMP.2. The loaded particles incorporate labelled ADP or ATP up to 6 pmol/g protein relatively 3. By measuring and labelling the endogenous nucleotide pool, the nucleotide transport was identified as a counter-exchange. 4.In contrast to mitochondria the inhibition by bongkrekate is rapid, independent of pH and preincubation with ADP. This indicates that bongkrekate inhibits the carrier from the outside.5. The exchange is inhibited by bongkrekate but not by atractylate unless the particles are derived from mitochondria preloaded with atractylate. This indicates asymmetry of the translocation with respect to its orientation to the membrane side: When oriented to the 'm-side' (outside in submitochondrial particles) the affinity is high for bongkrekate and low for atractylate and in the 'side' it is high for atractylate and low for bongkrekate. These results are in accordance with the predictions made from studies on mitochondria. In both states it has high affinity for palmityl-CoA.6. The translocation rate in sonic particles is about 18 pmol x min-l x g protein-' at 20 "C and amounts to 6 % of the rate in the parent mitochondria. Translocation rate has a high temperature dependence ((Ilo = 3.5) with E, = 24 kcal (100 kJ) in the range between 0 and 24 "C and between 24 "C and 35 "C, E, = 4 kcal (17 kJ) 'break' in the temperature dependence is at about 25 "C.7. Specificity of the nucleotide exchange is even higher than that of mitochondria. Exchange with dADP and AMP-P(NH)P reaches only about 3 % of the activity with ADP. The ADP uptake rate is about 30 % faster than that of ATP. There is little influence of uncoupling by carbonylcyanide p-trifluoromethoxyphenylhydrazone in contrast to mitochondria. 8. The exchange is not inhibited by N-ethylmaleimide unless N-ethylmaleimide is added prior to the sonication.The results demonstrate that in sonic particles the nucleotide can be transported similar to that in mitochondria, however, with strongly reduced activity. The exposure of the m-side in the particles gives important results about the asymmetry of the carrier properties.slowly (10 "C, t 1 / 2 = 20 s).The studies on the ADP/ATP exchange in mitochondria are extended to submitochondrial particles as derived from mitochondria by sonication. Submitochondrial particles represent a simpler preparation of the inner mitochondrial membrane, devoid of the outer membrane and of matrix proteins.Abbreviations. ANP, adenine nucleotide; m-side, the inner side of the mitochondrial membrane which is outside in the sonicated particles ; c-side, inner side ...

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“…A more penetrating interpretation encompasing the various effects of ADP, uncoupler, inhibitors on the NEM interaction with the carrier could be derived from our reorienting site mechanism of the ADP, ATP carrier. This interpretation is shown in the reaction scheme of fig.3 which represents the reaction step of the reorienting mechanism [7,8] modified to include the interaction of phenylglyoxal. Only a few of the experimental arguments for this scheme can be mentioned here.…”

Section: Resultsmentioning

confidence: 99%

Is there a common binding center in the ADP, ATP carrier for substrate and inhibitors?

Klingenberg

Appel

1980

FEBS Letters

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Experimental Demonstration of the Reorienting (Mobile) Carrier Mechanism Exemplified by the Mitochondrial Adenine Nucleotide Translocator

Cited by 27 publications

References 22 publications

The new atractyloside type compound as a high affinity ligand to the adenine nucleotide carrier

The new atractyloside type compound as a high affinity ligand to the adenine nucleotide carrier

The Adenine-Nucleotide Exchange in Submitochondrial (Sonic) Particles

Is there a common binding center in the ADP, ATP carrier for substrate and inhibitors?

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