BIOMAT Database Logo BIOMAT Database Logo

Photoaffinity labeling of the mitochondrial phosphate carrier by 4-azido-2-nitrophenyl phosphate. 1987

M Tommasino, and G Prezioso, and F Palmieri

The effect of 4-azido-2-nitrophenyl phosphate (ANPP), a photoreactive analogue of phosphate, on the phosphate carrier of pig-heart mitochondria has been investigated. In the dark, ANPP inhibits the transport of phosphate in a competitive manner with a Ki of 3.2 mM. Upon photoirradiation with visible light, [32P]ANPP binds covalently to the phosphate carrier and the inhibition becomes irreversible. Both the inhibition of phosphate transport and the incorporation of [32P]ANPP into the phosphate carrier depend on the concentration of the inhibitor and the pH of the medium. Incubation of the mitochondria with phosphate during illumination in the presence of ANPP protects the carrier against inactivation and decreases the amount of radioactivity which is found to be associated with the purified protein. By extrapolation it is calculated that at 100% inactivation of the phosphate carrier 0.35 mol of reagent are bound per mol of 33 kDa carrier protein. It is concluded that ANPP can be used for photoaffinity labeling of the mitochondrial phosphate carrier at the substrate-binding site.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008027 Light That portion of the electromagnetic spectrum in the visible, ultraviolet, and infrared range. Light, Visible,Photoradiation,Radiation, Visible,Visible Radiation,Photoradiations,Radiations, Visible,Visible Light,Visible Radiations
D008929 Mitochondria, Heart The mitochondria of the myocardium. Heart Mitochondria,Myocardial Mitochondria,Mitochondrion, Heart,Heart Mitochondrion,Mitochondria, Myocardial
D008933 Mitochondrial Swelling An increase in MITOCHONDRIAL VOLUME due to an influx of fluid; it occurs in hypotonic solutions due to osmotic pressure and in isotonic solutions as a result of altered permeability of the membranes of respiring mitochondria. Giant Mitochondria,Megamitochondria,Mitochondrial Hypertrophy,Giant Mitochondrias,Hypertrophy, Mitochondrial,Megamitochondrias,Mitochondria, Giant,Mitochondrial Hypertrophies,Swelling, Mitochondrial
D002352 Carrier Proteins Proteins that bind or transport specific substances in the blood, within the cell, or across cell membranes. Binding Proteins,Carrier Protein,Transport Protein,Transport Proteins,Binding Protein,Protein, Carrier,Proteins, Carrier
D003624 Darkness The absence of light. Darknesses
D000345 Affinity Labels Analogs of those substrates or compounds which bind naturally at the active sites of proteins, enzymes, antibodies, steroids, or physiological receptors. These analogs form a stable covalent bond at the binding site, thereby acting as inhibitors of the proteins or steroids. Affinity Labeling Reagents,Labeling Reagents, Affinity,Labels, Affinity,Reagents, Affinity Labeling
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001386 Azides Organic or inorganic compounds that contain the -N3 group. Azide
D012969 Sodium Fluoride A source of inorganic fluoride which is used topically to prevent dental caries. Fluoristat,Ossin,Zymafluor,Fluoride, Sodium,Fluorides, Sodium,Fluoristats,Ossins,Sodium Fluorides,Zymafluors

Related Publications

M Tommasino, and G Prezioso, and F Palmieri
Photoaffinity labeling of the mitochondrial oxoglutarate carrier by azido-phthalonate.
March 1995, Biochimica et biophysica acta,
M Tommasino, and G Prezioso, and F Palmieri
Photoaffinity labeling of the active site of the Na+/K(+)-ATPase with 4-azido-2-nitrophenyl phosphate.
January 1996, Biochemistry,
M Tommasino, and G Prezioso, and F Palmieri
Photoaffinity labeling of mitochondrial proteins with 2-azido [32P]palmitoyl CoA.
May 1995, FEBS letters,
M Tommasino, and G Prezioso, and F Palmieri
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 triphosphatase.
September 1980, Biochemistry,
M Tommasino, and G Prezioso, and F Palmieri
Photoaffinity labeling of peptide binding sites of prolyl 4-hydroxylase with N-(4-azido-2-nitrophenyl)glycyl-(Pro-Pro-Gly)5.
November 1985, Biochemistry,
M Tommasino, and G Prezioso, and F Palmieri
Identification of an active site peptide of skeletal myosin after photoaffinity labeling with N-(4-azido-2-nitrophenyl)-2-aminoethyl diphosphate.
March 1985, Proceedings of the National Academy of Sciences of the United States of America,
M Tommasino, and G Prezioso, and F Palmieri
Characterization of a phosphate binding domain on the alpha-subunit of chloroplast ATP synthase using the photoaffinity phosphate analogue 4-azido-2-nitrophenyl phosphate.
November 2000, Biochemistry,
M Tommasino, and G Prezioso, and F Palmieri
Photoaffinity labeling of rat liver carbamoyl phosphate synthetase I by 8-azido-ATP.
July 1987, The Journal of biological chemistry,
M Tommasino, and G Prezioso, and F Palmieri
Labeling of lymphocyte surface antigens by the lipophilic, photoactivatable reagent hexanoyldiiodo-N-(4-azido-2-nitrophenyl)tyramine.
June 1980, Biochemistry,
M Tommasino, and G Prezioso, and F Palmieri
Azido derivatives of dicarboxylic acids for photoaffinity labeling of mitochondrial carriers.
March 1989, Journal of biochemical and biophysical methods,
Copied contents to your clipboard!