Differentiation between mutants of Escherichia coli K12 defective in oxidative phosphorylation

Baruch I. Kanner, Nathan Nelson, David L. Gutnick*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


Hybrid membrane particles from two mutants of Escherichia coli K12, Bv4 and KI1, defective in oxidative phosphorylation, have been prepared, in which ATP-driven membrane energization is restored. A soluble factor of mutant KI1 was found to have properties similar to parental crude coupling factor, ATPase (EC Membrane particles of this mutant could not be reconstituted by parental coupling factor. Either parental coupling factor, or the soluble factor of mutant KI1 could reconstitute both respiration-driven and ATP-driven energization to membrane particles of mutant BV4 or to parental particles depleted of ATPase. Mutant BV4 was found to be devoid of coupling factor activity, while retaining the ability to hydrolyze ATP. Both mutants possess an ATPase with an altered binding to the membrane. Mutant KI1 is impaired in respiration-driven amino acid transport, in contrast to mutant BV4. The three major subunits of parental Escherichia coli ATPase have been isolated and antibodies have been prepared against these subunits. Antibodies against the largestsubunit (α component) or against the intact catalytic subunits (α + β components) inhibit both ATP-Pi exchange in the parent organism as well as ATP hydrolytic activity in parent and mutants. Antibodies against the two other subunits (β or γ components) also inhibit these two reactions, but were found to be less effective. Mutant NI44, which lacks ATPase activity, shows no precipitin lines with anti-α, anti-β, anti-γ, or anti-(α + β) preparations. In contrast, mutants BV4 and KI1, exhibit cross-reactivity with all of the antisera.

Original languageEnglish
Pages (from-to)347-359
Number of pages13
JournalBiochimica et Biophysica Acta - Bioenergetics
Issue number3
StatePublished - 8 Sep 1975


FundersFunder number
United States - Israel Binational Agricultural Research and Development Fund
United States-Israel Binational Science Foundation


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