Exploration of nucleotide binding sites in the mitochondrial membrane by 2-azido-[α-32 P]ADP
- 28 January 1985
- journal article
- Published by Wiley in FEBS Letters
- Vol. 180 (2) , 212-218
- https://doi.org/10.1016/0014-5793(85)81073-5
Abstract
The ADP/ATP carrier of beef heart mitochondria is able to bind 2‐azido‐[α‐32P]ADP in the dark with a K d value of ∼ 8 μM. 2‐Azido ADP is not transported and it inhibits ADP transport and ADP binding. Photoirradiation of beef heart mitochondria with 2‐azido‐[α‐32P]ADP results mainly in photolabeling of the ADP/ATP carrier protein; photolabeling is prevented by carboxyatractyloside, a specific inhibitor of ADP/ATP transport. Upon photoirradiation of inside‐out submitochondrial particles with 2‐azido‐[α‐32P]ADP, both the ADP/ATP carrier and the β subunit of the membrane‐bound F1‐ATPase are covalently labeled. The binding specificity of 2‐azido‐[α‐32P]ADP for the β subunit of F1‐ATPase is ascertained by prevention of photolabeling of isolated F1 by preincubation with an excess of ADP.Keywords
This publication has 16 references indexed in Scilit:
- Photoaffinity labeling with 2-azidoadenosine diphosphate of a tight nucleotide binding site on chloroplast coupling factor 1Proceedings of the National Academy of Sciences, 1982
- Binding of 2-azidoadenosine [.beta.-32P]diphosphate to the receptor on intact human blood platelets which inhibits adenylate cyclaseBiochemistry, 1982
- Adenine nucleotide transport in sonic submitochondrial particles. Kinetic properties and binding of specific inhibitorsBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1977
- Nuclear magnetic resonance studies of 5'-ribo- and deoxyribonucleotide structures in solutionBiochemistry, 1974
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- [13] Preparation, properties, and conditions for assay of mitochondria: Slaughterhouse material, small-scalePublished by Elsevier ,1967
- A rapid and improved method for the synthesis of 32P-labelled ribonucleoside 5′-monophosphatesBiochemical and Biophysical Research Communications, 1966
- Studies on the Azidoazomethine—Tetrazole Equilibrium. IV. Azidopurines1The Journal of Organic Chemistry, 1966
- Conversion of Mono- and Oligodeoxyribonucleotides to 5'-Triphosphates1Journal of the American Chemical Society, 1965
- Synthesis of Potential Anticancer Agents. XIV. Ribosides of 2,6-Disubstituted PurinesJournal of the American Chemical Society, 1958