Stability of the Escherichia coli ATP synthase F0F1 complex is dependent on interactions between γGln-269 and the β subunit loop βAsp- 301-βAsp-305

Hiroshi Omote, Ken Ichi Tainaka, Kazunari Fujie, Atsuko Iwamoto-Kihara, Yoh Wada, Masamitsu Futai

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The role of the conserved sequence motif 301DDL-TDP306 in the F0F1 ATP synthase β subunit was assessed by mutagenic analysis in the Escherichia coli enzyme. Mutations gave variable effects on F1 sector activity, stability, and membrane binding to the F0 sector. Upon solubilization, F1 sectors of the βD302E and βD305E mutants (βAsp-302 and βAsp-305 replaced by glutamate) dissociated into subunits, while mutants with other β305 substitutions failed to assemble. Membrane ATPase activities of β301 and 302 mutants were 20-70% of wild type. Replacements of the γ subunit Gln-269 had similar effects. The membrane ATPase activities of the γQ269E or γQ269D mutants were significantly lower and their F1 sectors dissociated into subunits upon solubilization. These results suggest that the β301-305 loop and the γ subunit region around Gln-269 form a key region for the assembly of α3β3γ complex. These results are consistent with the X- ray crystallographic structure of bovine F1 (J.P. Abrahams, A. G. W. Leslie, R. Lutter, and J. E. Walker (1994) Nature 370, 621-628) where the β301DDLTD305 loop directly interacts with γGln-269.

Original languageEnglish
Pages (from-to)277-282
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume358
Issue number2
DOIs
Publication statusPublished - Oct 15 1998
Externally publishedYes

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Escherichia coli
Adenosine Triphosphate
Membranes
Adenosine Triphosphatases
Conserved Sequence
Glutamic Acid
Substitution reactions
X-Rays
X rays
Mutation
Enzymes

Keywords

  • Assembly
  • F-ATPase

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Stability of the Escherichia coli ATP synthase F0F1 complex is dependent on interactions between γGln-269 and the β subunit loop βAsp- 301-βAsp-305. / Omote, Hiroshi; Tainaka, Ken Ichi; Fujie, Kazunari; Iwamoto-Kihara, Atsuko; Wada, Yoh; Futai, Masamitsu.

In: Archives of Biochemistry and Biophysics, Vol. 358, No. 2, 15.10.1998, p. 277-282.

Research output: Contribution to journalArticle

Omote, Hiroshi ; Tainaka, Ken Ichi ; Fujie, Kazunari ; Iwamoto-Kihara, Atsuko ; Wada, Yoh ; Futai, Masamitsu. / Stability of the Escherichia coli ATP synthase F0F1 complex is dependent on interactions between γGln-269 and the β subunit loop βAsp- 301-βAsp-305. In: Archives of Biochemistry and Biophysics. 1998 ; Vol. 358, No. 2. pp. 277-282.
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abstract = "The role of the conserved sequence motif 301DDL-TDP306 in the F0F1 ATP synthase β subunit was assessed by mutagenic analysis in the Escherichia coli enzyme. Mutations gave variable effects on F1 sector activity, stability, and membrane binding to the F0 sector. Upon solubilization, F1 sectors of the βD302E and βD305E mutants (βAsp-302 and βAsp-305 replaced by glutamate) dissociated into subunits, while mutants with other β305 substitutions failed to assemble. Membrane ATPase activities of β301 and 302 mutants were 20-70{\%} of wild type. Replacements of the γ subunit Gln-269 had similar effects. The membrane ATPase activities of the γQ269E or γQ269D mutants were significantly lower and their F1 sectors dissociated into subunits upon solubilization. These results suggest that the β301-305 loop and the γ subunit region around Gln-269 form a key region for the assembly of α3β3γ complex. These results are consistent with the X- ray crystallographic structure of bovine F1 (J.P. Abrahams, A. G. W. Leslie, R. Lutter, and J. E. Walker (1994) Nature 370, 621-628) where the β301DDLTD305 loop directly interacts with γGln-269.",
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