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N‐terminal acetylation of ectopic recombinant proteins in Escherichia coli
Author(s) -
Charbaut Elodie,
Redeker Virginie,
Rossier Jean,
Sobel André
Publication year - 2002
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/s0014-5793(02)03421-x
Subject(s) - acetylation , escherichia coli , recombinant dna , mass spectrometry , stathmin , tandem mass spectrometry , chemistry , biochemistry , biology , microbiology and biotechnology , chromatography , gene
N‐terminal acetylation is a protein modification common in eukaryotes, but rare in prokaryotes. Here, we characterized five mammalian stathmin‐like subdomains expressed in Escherichia coli by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry and nanoESI Q‐TOF tandem mass spectrometry. We revealed that RB3 SLD and RB3′ SLD are N α ‐acetylated, whereas SCG10 SLD and SCLIP SLD , although identical up to residue 6, are not, as well as stathmin. To assess the influence of the N‐terminal sequences on N α ‐acetylation, we exchanged residues 7 and 8 between acetylated RB3 SLD and unacetylated SCG10 SLD , and showed that it reversed the acetylation pattern. Our results demonstrate that ectopic recombinant proteins can be extensively N α ‐acetylated in E. coli , and that the rules governing N α ‐acetylation are complex and involve the N‐terminal region, as in eukaryotes.