Transmembrane topology of Escherichia coli H+-ATPase (ATP synthase) subunit a

Hiroshi Yamada, Yoshinori Moriyama, Masatomo Maeda, Masamitsu Futai

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Escherichia coli H+-ATPase subunit a is a hydrophobic F0 subunit. To investigate the topology of the subunit in the membrane, we prepared site-specific polyclonal antibodies against amino-terminal (Ser-3 to Leu-16), middle loop (Lys-167 to Gln-181), and carboxyl-terminal (Thr-259 to His-271) peptide segments. Enzyme-linked immunosorbent assay revealed that these antibodies specifically reacted with subunit a of inside-out membrane vesicles, but not with that of right-side-out spheroplasts. Full reactivity appeared when spheroplasts were disrupted with Triton X-100 (0.5%) or by sonication. These results suggest that at least parts of the three peptide segments of subunit a face the cytoplasm. Based on these observations, we propose a novel transmembrane topology of subunit a.

Original languageEnglish
Pages (from-to)34-38
Number of pages5
JournalFEBS Letters
Volume390
Issue number1
DOIs
Publication statusPublished - Jul 15 1996
Externally publishedYes

Fingerprint

Spheroplasts
Proton-Translocating ATPases
Escherichia coli
Adenosine Triphosphate
Topology
Membranes
Immunosorbents
Peptides
Sonication
Antibodies
Octoxynol
Assays
Cytoplasm
Enzyme-Linked Immunosorbent Assay
Enzymes

Keywords

  • ATP synthase
  • H-ATPase
  • Hydrophobic F subunit
  • Subunit a

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Transmembrane topology of Escherichia coli H+-ATPase (ATP synthase) subunit a. / Yamada, Hiroshi; Moriyama, Yoshinori; Maeda, Masatomo; Futai, Masamitsu.

In: FEBS Letters, Vol. 390, No. 1, 15.07.1996, p. 34-38.

Research output: Contribution to journalArticle

Yamada, Hiroshi ; Moriyama, Yoshinori ; Maeda, Masatomo ; Futai, Masamitsu. / Transmembrane topology of Escherichia coli H+-ATPase (ATP synthase) subunit a. In: FEBS Letters. 1996 ; Vol. 390, No. 1. pp. 34-38.
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