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

FEBS Letters

Futai, Masamitsu¹  Maeda, Masatomo¹  Moriyama, Yoshinori¹  Yamada, Hiroshi¹ 

[1] Department of Biological Science, The Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka 567, Japan

关键词: H+-ATPase;    ATP synthase;    Subunit a;    Hydrophobic F0 subunit;    PBS;    phosphate-buffered saline;    ELISA;    enzyme-linked immunosorbent assay;    F0F1;    Escherichia coli F0F1-ATP synthase;   

DOI  :  10.1016/0014-5793(96)00621-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown