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. 1974 Jul;71(7):2725–2729. doi: 10.1073/pnas.71.7.2725

Purification and Properties of Reconstitutively Active and Inactive Adenosinetriphosphatase from Escherichia coli

Masamitsu Futai 1, Paul C Sternweis 1, Leon A Heppel 1
PMCID: PMC388541  PMID: 4153028

Abstract

The Mg2+- and Ca2+-stimulated ATPase (EC 3.6.1.3; ATP phosphohydrolase) (bacterial coupling factor) was purified from two strains of E. coli by two different procedures: (a) method of Nelson, Kanner, and Gutnick [Proc. Nat. Acad. Sci. USA (1974) 71, 2720-2724] and (b) a modified procedure described in this paper. The ATPase purified from E. coli K12 (λ) by the first procedure had 4 subunits (α, β, γ, and ε). It did not bind to a deficient membrane, nor did it reconstitute ATP-driven transhydrogenase activity. Our modified procedure (b) yielded 5 subunits (α, β, γ, δ, and ε). This ATPase could bind to a deficient membrane and reconstitute ATP-driven transhydrogenase. This finding suggests that the δ subunit is required for the reaction with the membrane. The molecular weight of the 4-subunit ATPase was significantly lower than that of the 5-subunit ATPase, as judged by equilibrium centrifugation. The specific ATPase activities of both preparations were about the same. These two procedures were also applied to E. coli ML308-225.

Keywords: coupling factor, ATP-driven transhydrogenase

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Selected References

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