Site-Specific N- and C-Terminal Labeling of a Single Polypeptide Using Sortases of Different Specificity

被引:181
作者
Antos, John M. [1 ]
Chew, Guo-Liang [1 ]
Guimaraes, Carla P. [1 ]
Yoder, Nicholas C. [1 ]
Grotenbreg, Gijsbert M. [1 ]
Popp, Maximilian Wei-Lin [1 ]
Ploegh, Hidde L. [1 ]
机构
[1] Whitehead Inst Biomed Res, Cambridge, MA 02142 USA
基金
美国国家卫生研究院;
关键词
SURFACE PROTEIN;
D O I
10.1021/ja902681k
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
The unique reactivity of two sortase enzymes, SrtA(staph) from Staphylococcus aureus and SrtA(strep) from Streptococcus pyogenes, is exploited for site-specific labeling of a single polypeptide with different labels at its N and C termini. SrtA(strep) is used to label the protein's C terminus at an LPXTG site with a fluorescently labeled dialanine nucleophile. Selective N-terminal. labeling of proteins containing N-terminal glycine residues is achieved using SrtA(staph) and LPXT derivatives. The generality of N-terminal labeling with SrtA(staph) is demonstrated by near-quantitative labeling of multiple protein substrates with excellent site specificity.
引用
收藏
页码:10800 / +
页数:3
相关论文
共 9 条
  • [1] Analysis of the substrate specificity of the Staphylococcus aureus sortase transpeptidase SrtA
    Kruger, RG
    Otvos, B
    Frankel, BA
    Bentley, M
    Dostal, P
    McCafferty, DG
    [J]. BIOCHEMISTRY, 2004, 43 (06) : 1541 - 1551
  • [2] Surface protein IsdC and sortase B are required for heme-iron scavenging of Bacillus anthracis
    Maresso, Anthony W.
    Chapa, Travis J.
    Schneewind, Olaf
    [J]. JOURNAL OF BACTERIOLOGY, 2006, 188 (23) : 8145 - 8152
  • [3] Sortases and the art of anchoring proteins to the envelopes of gram-positive bacteria
    Marraffini, LA
    DeDent, AC
    Schneewind, O
    [J]. MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, 2006, 70 (01) : 192 - +
  • [4] An iron-regulated sortase anchors a class of surface protein during Staphylococcus aureus pathogenesis
    Mazmanian, SK
    Ton-That, H
    Su, K
    Schneewind, O
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (04) : 2293 - 2298
  • [5] An embarrassment of sortases - a richness of substrates?
    Pallen, MJ
    Lam, AC
    Antonio, M
    Dunbar, K
    [J]. TRENDS IN MICROBIOLOGY, 2001, 9 (03) : 97 - 101
  • [6] Sortagging: a versatile method for protein labeling
    Popp, Maximilian W.
    Antos, John M.
    Grotenbreg, Gijsbert M.
    Spooner, Eric
    Ploegh, Hidde L.
    [J]. NATURE CHEMICAL BIOLOGY, 2007, 3 (11) : 707 - 708
  • [7] Crystal Structure of Streptococcus pyogenes Sortase A IMPLICATIONS FOR SORTASE MECHANISM
    Race, Paul R.
    Bentley, Matthew L.
    Melvin, Jeff A.
    Crow, Allister
    Hughes, Richard K.
    Smith, Wendy D.
    Sessions, Richard B.
    Kehoe, Michael A.
    McCafferty, Dewey G.
    Banfield, Mark J.
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2009, 284 (11) : 6924 - 6933
  • [8] Sortase-Mediated Ligation: A Gift from Gram-Positive Bacteria to Protein Engineering
    Tsukiji, Shinya
    Nagamune, Teruyuki
    [J]. CHEMBIOCHEM, 2009, 10 (05) : 787 - 798
  • [9] Expansion of the sortase-mediated labeling method for site-specific N-terminal labeling of cell surface proteins on living cells
    Yamamoto, Teruyasu
    Nagamune, Teruyuki
    [J]. CHEMICAL COMMUNICATIONS, 2009, (09) : 1022 - 1024