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The in vivo assembly and function of the N‐ and C‐terminal halves of the Tn 10‐encoded TetA protein in Escherichia coli
Author(s) -
Yamaguchi Akihito,
Someya Yuichi,
Sawai Tetsuo
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)81378-d
Subject(s) - escherichia coli , terminal (telecommunication) , in vivo , escherichia coli proteins , function (biology) , chemistry , microbiology and biotechnology , biology , biochemistry , genetics , gene , computer science , telecommunications
The tet A gene was cut into its N‐ and C‐terminal halves at the central Eco RI site and the two halves were subcloned individually or together under a separate lac promoter/operator. The expression of the C‐terminal half was detected with a C‐terminal‐specific antibody. The amount of the N‐terminal half in the cytoplasmic membrane was not affected by the presence of the C‐terminal half. In contrast, the amount of the C‐terminal half in the membrane was increased in the presence of the N‐terminal half, indicating that the N‐terminal half helps the stable folding of the C‐terminal half in the membrane. Each half individually showed no tetracycline transport activity, however, when both halves were expressed together, the resultant complex showed about 40% of the tetracycline transport activity of the wild‐type per number of the C‐terminals of TetA protein in the membrane.