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Photolabeling of the phosphate binding site of mitochondrial F1-ATPase by (/sup 32/P)azidonitrophenyl phosphate. Identification of the photolabeled amino acid residues

Journal Article · · Biochemistry; (United States)
OSTI ID:5723618
; ; ; ; ; ;
(/sup 32/P)Azidonitrophenyl phosphate ((/sup 32/P)ANPP) is a photoactivatable analogue of Pi. It competes efficiently with Pi for binding to the F1 sector of beef heart mitochondrial ATPase and photolabels the Pi binding site located in the beta subunit of F1. By cleavage of the photolabeled beta subunit of F1 with cyanogen bromide, trypsin, and chymotrypsin, bound (/sup 32/P)ANPP was localized in a fragment spanning Thr 299-Phe 326. By Edman degradation of the radiolabeled tryptic peptide spanning Ile 296-Arg 337, (/sup 32/P)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 (/sup 32/P)ANPP interacts with a glycine-rich sequence of the beta subunit, spanning Gly 156-Lys 162, which is spatially close to the photolabeled Ile 304-Tyr 311 segment of the same subunit.
Research Organization:
Centre d'Etudes Nucleaires, Grenoble (France)
OSTI ID:
5723618
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 28:4; ISSN BICHA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACID ANHYDRASES
AFFINITY
AMINO ACID SEQUENCE
ANIMALS
ATP-ASE
AZIDES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CARDIOVASCULAR SYSTEM
CATTLE
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
ENZYMES
FLUORESCENCE
HEART
HYDROLASES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLING
LIGHT NUCLEI
LUMINESCENCE
MAMMALS
MEMBRANE PROTEINS
MITOCHONDRIA
MOLECULAR STRUCTURE
NITROGEN COMPOUNDS
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANOIDS
ORGANS
OXYGEN COMPOUNDS
PHOSPHATES
PHOSPHOHYDROLASES
PHOSPHORUS 32
PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
RUMINANTS
TRACER TECHNIQUES
VERTEBRATES