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Expression of the human UDP‐glucuronosyltransferase UGT1 1 6 in Escherichia coli
Author(s) -
Ouzzine Mohamed,
Fournel-Gigleux Sylvie,
Pillot Thierry,
Burchell Brian,
Siest Gérard,
Magdalou Jacques
Publication year - 1994
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(94)80414-1
Subject(s) - signal peptide , escherichia coli , chemistry , biochemistry , recombinant dna , peptide , mutagenesis , peptide sequence , gene , mutation
The membrane‐bound human liver UDP‐glucuronosyltransferase UGT1 ∗ 6 was expressed in Escherichia coli . Exchange of the natural signal peptide by the bacterial signal peptides of pelB or OmpT proteins considerably increased the level of expression and, as the natural signal peptide, targeted the protein to the membranes. The extent of maturation of s pelB‐UGTl*6 precursor was about 30%. No processing of s OmpT‐UGTl ∗ 6 occurred but the processing rate of this precursor could be significantly increased by mutagenesis of the first two amino acid residues of the mature sequence. These expression vectors allowed us to produce high levels of recombinant mature UGT1 ∗ 6 required for further structural studies.