Photolabeling of the phosphate binding site of mitochondrial F1-ATPase by [32P]azidonitrophenyl phosphate. Identification of the photolabeled amino acid residues
- 21 February 1989
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 28 (4) , 1442-1448
- https://doi.org/10.1021/bi00430a003
Abstract
[32P]Azidonitrophenyl phosphate ([32P]ANPP) is a photoactivatable analogue of Pi. It competes efficiently with Pi for binding to the F1 sector of beef heart mitochondrial ATPase and photolables the Pi binding site located in the .beta. subunit of F1 [Lauquin, G. J. M., Pougeois, R., and Vignais, P. V. (1980) Biochemistry 19, 4620-4626]. By cleavage of the photolabeled .beta. subunit of F1 with cyanogen bromide, trypsin, and chymotrypsin, bound [32P]ANPP was localized in a fragment spanning Thr 299-Phe 326. By Edman degradation of the radiolabeled tryptic peptide spanning Ile 296-Arg 337, [32P]ANPP was found to be attached covalently by its photoreactive group to Ile 304, Gln 308, and Tyr 311. These results are discussed in terms of a model in which the phosphate group of [32P]ANPP interacts with a glycine-rich sequence of the .beta. subunit, spannig Gly 156-Lys 162, which is spatially close to the photolabeled Ile 304-Tyr 311 segment of the same subunit.This publication has 35 references indexed in Scilit:
- Photoaffinity labeling of the mitochondrial phosphate carrier by 4-azido-2-nitrophenyl phosphateBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1987
- Identification of amino acid residues photolabeled with 2-azido[.alpha.-32P]adenosine diphosphate in the .beta. subunit of beef heart mitochondrial F1-ATPaseBiochemistry, 1986
- Labeling of individual amino acid residues in the membrane‐embedded F0 part of the F1 F0 ATP synthase from Neurospora crassaEuropean Journal of Biochemistry, 1986
- A Model for the Tertiary Structure of p21, the Product of the ras OncogeneScience, 1985
- Tyrosine-311 of a beta chain is the essential residue specifically modified by 4-chloro-7-nitrobenzofurazan in bovine heart mitochondrial ATPaseEuropean Journal of Biochemistry, 1985
- Aurovertin binding sites on beef heart mitochondrial F1-ATPase. Study with [14C]aurovertin D of the binding stoichiometry and of the interaction between aurovertin and the natural ATPase inhibitor for binding to F1Biochemistry, 1983
- Interaction of 4-azido-2-nitrophenyl phosphate, an inorganic phosphate photoreactive analog, with chloroplast coupling factor 1Biochemistry, 1983
- Spontaneous aggregation of the mitochondrial natural ATPase inhibitor in salt solutions as demonstrated by gel filtration and neutron scattering. Application to the concomitant purification of the ATPase inhibitor and F1-ATPaseBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1982
- Effect of the natural ATPase inhibitor on the binding of adenine nucleotides and inorganic phosphate to mitochondrial F1-ATPaseBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1981
- 4-Azido-2-nitrophenyl phosphate, a new photoaffinity derivative of inorganic phosphate. Study of its interaction with the inorganic phosphate binding site of beef heart mitochondrial adenosine triphosphataseBiochemistry, 1980