ATP-Pi Exchange Preparation from Escherichia Coli

N. Nelson, R. Chibovsky, D. L. Gutnick

Research output: Contribution to journalArticlepeer-review

Abstract

This chapter discusses the assay for the adenosine triphosphate–inorganic phosphate (ATP–Pi) exchange reaction in mitochondria, chloroplasts, and bacterial chromatophores. The basis of these investigations is the ability to detect incorporation of 32Pi into ATP mediated by the membrane ATPase under conditions approaching equilibrium. Because the bacterial cell exhibits a number of different phosphate transfer reactions between Pi and phosphonucleotides, it is necessary to differentiate between the reaction catalyzed by the membrane ATPase and other cellular activities. The exchange reaction associated with the energy-transducing ATPase can be distinguished by its sensitivity to uncoupling agents and energy-transfer inhibitors. The exchange activity should be inhibited by specific antibody prepared against pure Escherichia coli ATPase. The reaction is be observed in mutants of E. coli that are defective in oxidative phosphorylation by virtue of a genetic lesion in the membrane ATPase complex. The chapter describes some of the characteristics of the ATP–Pi exchange reaction in membrane particles isolated from the organism E. coli.

Original languageEnglish
Pages (from-to)358-363
Number of pages6
JournalMethods in Enzymology
Volume55
Issue numberC
DOIs
StatePublished - 1 Jan 1979

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