Orientation of the carboxyl terminus of the transposon Tn10‐encoded tetracycline resistance protein in Escherichia coli

FEBS Letters

Yamaguchi, Akihito¹  Adachi, Kumiko¹  Sawai, Tetsuo¹ 

[1] Division of Microbial Chemistry, Faculty of Pharmaceutical Sciences, Chiba University, Chiba 260, Japan

关键词: Tetracycline resistance protein;    Tetracycline;    Antiporter;    TetA;    Topology;    Site-specific antibody;    RSOV;    right-side-out vesicles;    ISOV;    inside-out vesicles;    ELISA;    enzyme-linked immunosorbent assay;   

DOI  :  10.1016/0014-5793(90)80872-G

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A site-directed antibody was generated against a synthetic polypeptide corresponding to the 14 amino acid residues of the carboxyl terminus of the Tn10 TetA protein. The antibody reacted preferentially with inside-out vesicles, rather than right-side-out vesicles, prepared from Escherichia coli cells harboring transposon Tn10. When inside-out vesicles were treated with trypsin, the TetA protein was completely digested in the vicinity of the carboxyl terminus, as judged on immunoblot analysis using the antibody. In contrast, when right-side-out vesicles were treated with trypsin, the TetA protein was hardly digested. These results indicate that the carboxyl terminus of TetA is exposed to the cytoplasmic side of the membrane.

【 授权许可】

Unknown