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Further investigations on the inorganic phosphate binding site of beef heart mitochondrial F1-ATPase

Journal Article · · Biochemistry; (United States)
DOI:https://doi.org/10.1021/bi00325a032· OSTI ID:5023437
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The possibility that 4-azido-2-nitrophenyl phosphate (ANPP), a photoreactive derivative of inorganic phosphate (P /sub i/ ), could mimic ATP was investigated. ANPP was hydrolyzed in the dark by sarcoplasmic reticulum Ca/sup 2 +/-ATPase in the presence of Ca/sup 2 +/ but not in the presence of ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. ANPP was not hydrolyzed by purified mitochondrial F1-ATPase; however, ADP and ATP protected F1-ATPase against ANPP photoinactivation. On the other hand, the trinitrophenyl nucleotide analogues (TNP-ADP, TNP-ATP, and TNP-AMP-PNP), which bind specifically at the two catalytic sites of F1-ATPase, abolished P /sub i/ binding on F1-ATPase; they do not protect F1-ATPase against ANPP photoinactivation. Furthermore, ANPP-photoinactivated F1-ATPase binds the TNP analogues in the same way as the native enzyme. The Pi binding site of F1-ATPase, which is shown to be photolabeled by ANPP, does not appear to be at the gamma-phosphate position of the catalytic sites.
OSTI ID:
5023437
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 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
ANIMALS
ATP
ATP-ASE
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CARDIOVASCULAR SYSTEM
CATTLE
CELL CONSTITUENTS
CHEMISTRY
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
ENZYME INHIBITORS
ENZYMES
HEART
HYDROLASES
INACTIVATION
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
MAMMALS
MITOCHONDRIA
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANOIDS
ORGANS
OXYGEN COMPOUNDS
PHOSPHATES
PHOSPHOHYDROLASES
PHOSPHORUS 32
PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
PHOTOCHEMISTRY
RADIOISOTOPES
REACTION KINETICS
RUMINANTS
SARCOPLASMIC RETICULUM
TRACER TECHNIQUES
VERTEBRATES