Participation of N
            <sub>1</sub>
            -Oxide derivatives of Adenine Nucleotides in the Phosphotransferase Activity of Liver Mitochondria

Jebeleanu, Gheorghe; Ty, N.G.; Mantsch, Henry H.; Bârzu, Octavian; Niac, Gavril; Abrudan, I.

doi:10.1073/pnas.71.11.4630

Cited by 9 publications

(8 citation statements)

References 27 publications

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“…The K,-value for ANP (41 pM) is lower but not significantly different from the Ki-value for rroANP (9.3 x loh5 M). Especially, these binding data indicate that the recognition between the carrier and the nucleotide is based primarily on the adenine heterocycle [8,28,29] and the three anionic charges of the phosphate chain [28] resulting in a specific binding of the nucleotide to the carrier. Nevertheless, these recognition elements are not sufficient to induce a nucleotide transfer across the mitochondrial membrane.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Properties of the Ribose‐Ring‐Opened Adenine Nucleotide, 2,2′[1‐(9‐adenyl)‐1′‐(tri‐,diphosphoryl‐oxymethyl)]‐dihydroxydiethylether in Mitochondrial Adenine‐Nucleotide Translocation

Boos

Schlimme

Bojanovski

et al. 1975

European Journal of Biochemistry

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(Adenine-'4C) or (y3'P)-labelled 2,2'[1-(9-adenyl)-I '-(tri-, diphosphoryl-oxymethy1)l-dihydroxydiethylether (rroANP) was obtained from ANP by cleavage of the C-2'-C-3' bond by sodium periodate oxidation and subsequent borohydride reduction.Binding of rroANP to rat liver mitochondria revealed carrier-linked (atractyloside-sensitive) and unspecific (atractyloside-insensitive) binding but no transfer across the inner mitochondrial membrane. Kinetic data indicate rroANP as acompetitive inhibitor for AhT uptake with Ki = 9.3 x M. Experimental rroANP confirmed that an intact adenine base and three anionic charges of the phosphate chain are essential for the recognition between AM'-carrier and nucleotide but insufficient for the induction of a transmembrane ANP exchange. In addition mobilisation of the carrier-nucleotide complex requires an intact ribofuranoside ring system. Mitochondria1 AM' translocation is a key step for energy supply processes of aerobic organisms. Much interest has been directed to the catalytic mechanism of this carrier-mediated adenine nucleotide transfer across the inner mitochondrial membrane in the last decade [1,2]. Up to now, the molecular nature of the membrane integral Am-carrier is still unknown, but lately isolation and characterisation techniques have been developed [3 -61. Studies of specific interactions between the carrier and its substrate are actually still confined to measurements in situ. Analogues of the adenine nucleotides, therefore, are very useful probes for studying structural and electronic factors involved in the catalytic mechanism of AAPdependent biological processes in general and, as shown in this paper, on mitochondrial ANP translocation [7,8, and cited references].This publication describes carrier-linked binding and exchange properties of the ribose-ring opened Am-analogue 2,2'[1-(9-adenyl)-l'-(tri-, diphosphoryl-oxymethyl)]-dihydroxydiethylether rro-AT(D)P. Our interest was directed to investigations of struc- tural requirements of the mitochondrial membrane integral Am-carrier for substrate binding and transfer across the inner mitochondrial membrane. EXPERIMENTAL PROCEDURE MATERIALS Preparation of rroATPThe ribose-ring opened ATP analogue was prepared by cleavage of the C-2' -C-3' bond in a modidure [9 -111 by sodium periodate oxidation and subsequent sodium borohydride reduction.8.25 pmol ATP disodium salt was dissolved in 0.22 ml 0.1 M phosphate buffer pH 7.0 and 0.44 ml 0.1 M sodium periodate were added. Excess periodate was destroyed by addition of 0.045 ml 1 M glucose solution. (Reaction time 45 min). After oxidation 5 m g sodium borohydride solved in 0.43ml double distilled water was added. The pH of the reaction mixture was adjusted to 7.8 by 2 M sodium acetate buffer pH 5.2. After a reaction period of 24 h at room temperature excess sodium borohydride was destroyed by adjusting the pH to 5.2. The reaction mixture was then applied to a DEAE-cellulose column (Whatman DEAE 32-cellulose, 1 x 5 cm). The modified ANP was eluted by a linear gradient of trieth...

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Section: Discussionmentioning

confidence: 99%

“…This shows again that binding properties are primarily related to an intact adenine base and the presence of three anionic charges in the phosphate chain [8,28]. For induction of carriermediated transmembrane ANP exchange, distinct conformational criteria must be fulfilled.…”

Section: Discussionmentioning

confidence: 99%

Properties of the Ribose‐Ring‐Opened Adenine Nucleotide, 2,2′[1‐(9‐adenyl)‐1′‐(tri‐,diphosphoryl‐oxymethyl)]‐dihydroxydiethylether in Mitochondrial Adenine‐Nucleotide Translocation

Boos

Schlimme

Bojanovski

et al. 1975

European Journal of Biochemistry

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“…While varying the 8-BrADP concentration, we kept the ATP concentration constant at 0.6 mM (O) or at 0.16 mM (•). The reaction was started with 10-20 µg of protein of partially purified mitochondrial NDP kinase [for details see Jebeleanu et al (1974)].…”

Section: Resultsmentioning

confidence: 99%

“…Oxidative phosphorylation, the ATP-linked reduction of NAD+ by succinate, and the ATPase activity of beef heart submitochondrial particles were assayed essentially as described by Penefsky (1974). The isolation of rat liver mitochondria, the measurement of mitochondrial respiration, and the exchange of intramitochondrial 14C-labeled adenine nucleotides with externally added nucleotide analogues were performed as described in previous publications (Jebeleanu et al, 1974;Mantsch et al, 1975). Rat kidney nucleoside diphosphate kinase was obtained according to Lascu and Bárzu (unpublished), pig kidney FDPase was obtained as described by Colombo et al (1972), rabbit muscle phosphoglycerate kinase was obtained according to Krietsch & Biicher (1970), and yeast pyruvate kinase was obtained as described by Yun et al (1976).…”

Section: Methodsmentioning

confidence: 99%

Enzymic properties of 8-bromoadenine nucleotides

Lascu¹,

Kezdi²,

Goia³

et al. 1979

Biochemistry

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8-Bromoadenine nucleotides were tested as potential substrates and/or inhibitors of mitochondrial processes in intact or disrupted organelles, as substrates of various phosphotransferases, and as allosteric effectors in the reactions catalyzed by phosphofructokinase, isocitrate dehydrogenase, glutamate dehydrogenase, and fructose-1,6-bisphosphatase. 8-BrATP and 8-BrADP are not recognized by the translocase system located in the inner mitochondrial membrane and cannot be used as usbstrates in oxidative phosphorylation and related reactions catalyzed be beef heart submitochondrial membranes. This confirms the high specificity for adenine nucleotides of the mammalian systems involved in energy-yielding and energy-requiring reactions. However, 8-BrATP and 8-BrADP are able to substitute for the natural adenine nucleotides in reactions catalyzed by many phosphotransferases, although their capacity as phosphate donors and acceptors is generally much reduced. On the other hand, in almost all investigated cases, the 8-bromoadenine nucleotides have lost the capability of the natural adenine nucleotides to act as allosteric effectors, indicating that the structural requirements for allosteric activity are more stringent than those for catalytic activity.

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“…Biochemical Assays. The isolation of rat liver mitochondria, the measurement of mitochondrial respiration, and the exchange of intramitochondrial 14C-labeled adenine nucleotides with externally added nucleotide analogues were performed as described in previous publications (Jebeleanu et al, 1974;Mantsch et al, 1975;Bárzu et al, 1976b). Heavy beef heart mitochondria were prepared as described by Low & Vallin (1963), stored for 2-7 days at -10 °C, and thawed only shortly before use.…”

Section: Methodsmentioning

confidence: 99%

Phosphorylation and hydrolysis of 7-deazaadenine nucleotides by rat liver and beef heart mitochondria

Petrescu¹,

Lascu²,

Goia³

et al. 1982

Biochemistry

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Tubercidin nucleotides [tubercidin 5'-mono-phosphate (TuMP), 5'-diphosphate (TuDP), and 5'-triphosphate (TuTP)] were tested as potential substrates for the mitochondrial phosphotransferases from rat liver and beef heart. TuDP is recognized by the mitochondrial translocase and phosphorylated by the respiratory chain enzymes in both mitochondria and submitochondrial particles from rat liver and beef heart; the low transport rate of the analogue into the matrix space of the intact organelles seems to be not a limiting step in the formation of TuTP. The phosphorylation of TuDP is significantly lower in beef heart mitochondria because of a higher specificity for ADP of the heart oxidative phosphorylation system. On the basis of the kinetic parameters of the partially purified liver mitochondrial adenylate kinase, one can conclude that the liver mitochondria are able to phosphorylate in vivo TuMP at a rate practically equal to the rate of AMP phosphorylation. The liver mitochondrial NDP kinase ensures a further fast phosphorylation of TuDP without the direct involvement of respiratory chain enzymes. In the case of heart mitochondria, two factors limit the rate of TuMP phosphorylation to TuTP: the lower acceptor activity of adenylate kinase with TuMP as compared with AMP and the different localization of heart NDP kinase situated on the inner face of the inner mitochondrial membrane. TuDP and TuTP preserve the ability of the natural nucleotides to interact with the "tight" nucleotide binding sites of isolated or membrane-bound F1. The low hydrolytic rate of TuTP with F1 may be related to the unusual flexibility of the glycosyl bond of tubercidin nucleotides in aqueous solution, with a high accessibility to syn conformation.

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Participation of N ₁ -Oxide derivatives of Adenine Nucleotides in the Phosphotransferase Activity of Liver Mitochondria

Abstract: The modified adenine nucleotides ATP-

Cited by 9 publications

References 27 publications

Properties of the Ribose‐Ring‐Opened Adenine Nucleotide, 2,2′[1‐(9‐adenyl)‐1′‐(tri‐,diphosphoryl‐oxymethyl)]‐dihydroxydiethylether in Mitochondrial Adenine‐Nucleotide Translocation

Properties of the Ribose‐Ring‐Opened Adenine Nucleotide, 2,2′[1‐(9‐adenyl)‐1′‐(tri‐,diphosphoryl‐oxymethyl)]‐dihydroxydiethylether in Mitochondrial Adenine‐Nucleotide Translocation

Enzymic properties of 8-bromoadenine nucleotides

Phosphorylation and hydrolysis of 7-deazaadenine nucleotides by rat liver and beef heart mitochondria

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Participation of N 1 -Oxide derivatives of Adenine Nucleotides in the Phosphotransferase Activity of Liver Mitochondria

Abstract: The modified adenine nucleotides ATP-

Cited by 9 publications

References 27 publications

Properties of the Ribose‐Ring‐Opened Adenine Nucleotide, 2,2′[1‐(9‐adenyl)‐1′‐(tri‐,diphosphoryl‐oxymethyl)]‐dihydroxydiethylether in Mitochondrial Adenine‐Nucleotide Translocation

Properties of the Ribose‐Ring‐Opened Adenine Nucleotide, 2,2′[1‐(9‐adenyl)‐1′‐(tri‐,diphosphoryl‐oxymethyl)]‐dihydroxydiethylether in Mitochondrial Adenine‐Nucleotide Translocation

Enzymic properties of 8-bromoadenine nucleotides

Phosphorylation and hydrolysis of 7-deazaadenine nucleotides by rat liver and beef heart mitochondria

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Participation of N ₁ -Oxide derivatives of Adenine Nucleotides in the Phosphotransferase Activity of Liver Mitochondria