Genetically encoding N(epsilon)-acetyllysine in recombinant proteins.
N(epsilon)-acetylation of lysine (1) is a reversible post-translational modification with a regulatory role that rivals that of phosphorylation in eukaryotes. No general methods exist to synthesize proteins containing N(epsilon)-acetyllysine (2) at defined sites. Here we demonstrate the site-specific incorporation of N(epsilon)-acetyllysine in recombinant proteins produced in Escherichia coli via the evolution of an orthogonal N(epsilon)-acetyllysyl-tRNA synthetase/tRNA(CUA) pair. This strategy should find wide applications in defining the cellular role of this modification.
Neumann
Heinz
H
Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 0QH England, UK.
Peak-Chew
Sew Y
SY
Chin
Jason W
JW
D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics; D017021:Methanosarcina barkeri / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008968:Molecular Conformation; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics*
D011994:Recombinant Proteins; C016949:N-epsilon-acetyllysine; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases; D008239:Lysine