Genetically encoding Nε-acetyllysine in recombinant proteins

被引：490

作者：

Neumann, Heinz ^{[1
]}

Peak-Chew, Sew Y. ^{[1
]}

Chin, Jason W. ^{[1
]}

机构：

[1] MRC, Lab Mol Biol, Cambridge CB2 0QH, England

来源：

NATURE CHEMICAL BIOLOGY | 2008年 / 4卷 / 04期

基金：

英国医学研究理事会;

关键词：

D O I：

10.1038/nchembio.73

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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/tRNACUA pair. This strategy should find wide applications in defining the cellular role of this modification.

引用

收藏

页码：232 / 234

页数：3

相关论文

共 15 条

[1] Pyrrolysine is not hardwired for cotranslational insertion at UAG codons [J].

Ambrogelly, Alexandre ;

Gundllapalli, Sarath ;

Herring, Stephanie ;

Polycarpo, Carla ;

Frauer, Carina ;

Soll, Dieter .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (09) :3141-3146

[2] Direct charging of tRNACUA with pyrrolysine in vitro and in vivo [J].

Blight, SK ;

Larue, RC ;

Mahapatra, A ;

Longstaff, DG ;

Chang, E ;

Zhao, G ;

Kang, PT ;

Church-Church, KB ;

Chan, MK ;

Krzycki, JA .

NATURE, 2004, 431 (7006) :333-335

[3] Addition of a photocrosslinking amino acid to the genetic code of Escherichia coli [J].

Chin, JW ;

Martin, AB ;

King, DS ;

Wang, L ;

Schultz, PG .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (17) :11020-11024

[4] A new UAG-encoded residue in the structure of a methanogen methyltransferase [J].

Hao, B ;

Gong, WM ;

Ferguson, TK ;

James, CM ;

Krzycki, JA ;

Chan, MK .

SCIENCE, 2002, 296 (5572) :1462-1466

[5] Translating the histone code [J].

Jenuwein, T ;

Allis, CD .

SCIENCE, 2001, 293 (5532) :1074-1080

[6] Structure of pyrrolysyl-tRNA synthetase, an archaeal enzyme for genetic code innovation [J].

Kavran, Jennifer M. ;

Gundliapalli, Sarath ;

O'Donoghue, Patrick ;

Englert, Markus ;

Soell, Dieter ;

Steitz, Thomas A. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (27) :11268-11273

[7] Substrate and functional diversity of lysine acetylation revealed by a proteomics survey [J].

Kim, Sung Chan ;

Sprung, Robert ;

Chen, Yue ;

Xu, Yingda ;

Ball, Haydn ;

Pei, Jimin ;

Cheng, Tzuling ;

Kho, Yoonjung ;

Xiao, Hao ;

Xiao, Lin ;

Grishin, Nick V. ;

White, Michael ;

Yang, Xiang-Jiao ;

Zhao, Yingming .

MOLECULAR CELL, 2006, 23 (04) :607-618

[8] Acetylation: a regulatory modification to rival phosphorylation? [J].

Kouzarides, T .

EMBO JOURNAL, 2000, 19 (06) :1176-1179

[9] Chromatin modifications and their function [J].

Kouzarides, Tony .

CELL, 2007, 128 (04) :693-705

[10] Pyrrolysine analogues as substrates for pyrrolysyl-tRNA synthetase [J].

Polycarpo, Carla R. ;

Herring, Stephanie ;

Berube, Amelie ;

Wood, John L. ;

Soll, Dieter ;

Ambrogelly, Alexandre .

FEBS LETTERS, 2006, 580 (28-29) :6695-6700