Amino acid activated adenosine triphosphate-pyrophosphate exchange by modified Echerichia coli cells

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Abstract

Amino acid activated adenosine 5′-triphosphate-[32P]pyrophosphate exchange has been studied using suspensions of Escherichia coli cells, and evidence is presented that this exchange reaction is carried out by unlysed cells. Exchange activity in minimal when freshly harvested cells are employed but increases after the cells have been frozen. Exchange activity is also markedly increased when the cells harvested during the logarithmic phase growth are incubated with ethylenediaminetetraacetate or polymyxin B before assay. Cells inhibited in the logarithmic phase of growth by streptomycin, chloramphenicol, or demethylchlorotetracycline show greater amino acid stimulated adenosine 5′-triphosphate-[32P]pyrophosphate exchange activity than do uninhibited cells. It is concluded that the enzymes which exchange adenosine 5′-triphosphate with [32P]pyrophosphate in the presence of amino acids can function at the surface of undisrupted E. coli cells and that the promotion of this activity may be a useful index of alteration of limiting membrane structures.

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Present address: Department of Pathology, School of Medicine, Yale University, New Haven, Conn. (U.S.A.).

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