N‐terminal acetylation of ectopic recombinant proteins in Escherichia coli

FEBS Letters

Redeker, Virginie¹  Rossier, Jean¹  Sobel, André²  Charbaut, Elodie² 

[1] CNRS UMR 7637, ESPCI, 10 rue Vauquelin, 75005 Paris, France;U440 INSERM/UPMC, Institut du Fer à Moulin, 17 rue du Fer à Moulin, 75005 Paris, France

关键词: N-terminal acetylation;    Mass spectrometry;    Ectopic protein expression;    Stathmin family;    Escherichia coli;    MS;    mass spectrometry;    MALDI-TOF;    matrix-assisted laser desorption ionization time-of-flight;    ESI;    electrospray ionization;    Q-TOF;    quadrupole time-of-flight;    TFA;    trifluoroacetic acid;    SLD;    stathmin-like domain;    m/z;    mass-to-charge ratio;   

DOI  :  10.1016/S0014-5793(02)03421-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown