Sortagging: a versatile method for protein labeling

被引：390

作者：

Popp, Maximilian W.

Antos, John M.

Grotenbreg, Gijsbert M.

Spooner, Eric

Ploegh, Hidde L.

机构：

[1] Whitehead Inst Biomed Res, Cambridge, MA 02142 USA

[2] MIT, Dept Biol, Cambridge, MA 02142 USA

来源：

NATURE CHEMICAL BIOLOGY | 2007年 / 3卷 / 11期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1038/nchembio.2007.31

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：

Genetically encoded reporter constructs that yield fluorescently labeled fusion proteins are a powerful tool for observing cell biological phenomena, but they have limitations. Sortagging (sortase-mediated transpeptidation) is a versatile chemoenzymatic system for site-specific labeling of proteins with small (< 2 kDa) probes. Sortagging combines the precision of a genetically encoded tag with the specificity of an enzymatic reaction and the ease and chemical versatility of peptide synthesis. Here we apply this technique to proteins in vitro and on the surface of living cells.

引用

收藏

页码：707 / 708

页数：2

相关论文

共 15 条

[1] Development of an HPLC assay for Staphylococcus aureus sortase:: Evidence for the formation of the kinetically competent acyl enzyme intermediate [J].

Aulabaugh, Ann ;

Ding, Weidong ;

Kapoor, Bhupesh ;

Tabei, Keiko ;

Alksne, Lefa ;

Dushin, Russ ;

Zatz, Tracy ;

Ellestad, George ;

Huang, Xinyi .

ANALYTICAL BIOCHEMISTRY, 2007, 360 (01) :14-22

[2] Site-specific labeling of proteins with small molecules in live cells [J].

Chen, I ;

Ting, AY .

CURRENT OPINION IN BIOTECHNOLOGY, 2005, 16 (01) :35-40

[3] Sorting sortases: a nomenclature proposal for the various sortases of Gram-positive bacteria [J].

Dramsi, S ;

Trieu-Cuot, P ;

Bierne, H .

RESEARCH IN MICROBIOLOGY, 2005, 156 (03) :289-297

[4] Multicolor and electron microscopic imaging of connexin trafficking [J].

Gaietta, G ;

Deerinck, TJ ;

Adams, SR ;

Bouwer, J ;

Tour, O ;

Laird, DW ;

Sosinsky, GE ;

Tsien, RY ;

Ellisman, MH .

SCIENCE, 2002, 296 (5567) :503-507

[5] Analysis of the substrate specificity of the Staphylococcus aureus sortase transpeptidase SrtA [J].

Kruger, RG ;

Otvos, B ;

Frankel, BA ;

Bentley, M ;

Dostal, P ;

McCafferty, DG .

BIOCHEMISTRY, 2004, 43 (06) :1541-1551

[6] Development and use of fluorescent protein markers in living cells [J].

Lippincott-Schwartz, J ;

Patterson, GH .

SCIENCE, 2003, 300 (5616) :87-91

[7] Overexpression and mislocalization of a tail-anchored GFP redefines the identity of peroxisomal ER [J].

Lisenbee, CS ;

Karnik, SK ;

Trelease, RN .

TRAFFIC, 2003, 4 (07) :491-501

[8] Sortase-mediated protein ligation: A new method for protein engineering [J].

Mao, HY ;

Hart, SA ;

Schink, A ;

Pollok, BA .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2004, 126 (09) :2670-2671

[9] Targeting proteins to the cell wall of sporulating Bacillus anthracis [J].

Marraffini, Luciano A. ;

Schneewind, Olaf .

MOLECULAR MICROBIOLOGY, 2006, 62 (05) :1402-1417

[10] Sortase A as a novel molecular "stapler" for sequence-specific protein conjugation [J].

Parthasarathy, Ranganath ;

Subramanian, Shyamsundar ;

Boder, Eric T. .

BIOCONJUGATE CHEMISTRY, 2007, 18 (02) :469-476