Orientation of the carboxyl terminus of the transposon Tn10-encoded tetracycline resistance protein in Escherichia coli.
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.
Yamaguchi
A
A
Division of Microbial Chemistry, Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Adachi
K
K
Sawai
T
T
D000595:Amino Acid Sequence; D000936:Antigen-Antibody Complex; D001426:Bacterial Proteins / Q000235:genetics; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D004251:DNA Transposable Elements; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis; D012097:Repressor Proteins / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D014157:Transcription Factors / Q000235:genetics*
D000936:Antigen-Antibody Complex; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D009842:Oligopeptides; D012097:Repressor Proteins; D014157:Transcription Factors; C034811:tetracycline resistance-encoding transposon repressor protein