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

John M. Antos | Zendy; Guo-Liang Chew | Zendy; Carla P. Guimarães | Zendy; Nicholas C. Yoder | Zendy; Gijsbert M. Grotenbreg | Zendy; Maximilian W. Popp | Zendy; Hidde L. Ploegh | Zendy

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

Author(s) -

John M. Antos,

Guo-Liang Chew,

Carla P. Guimarães,

Nicholas C. Yoder,

Gijsbert M. Grotenbreg,

Maximilian W. Popp,

Hidde L. Ploegh

Publication year - 2009

Publication title -

journal of the american chemical society

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 7.115

H-Index - 612

eISSN - 1520-5126

pISSN - 0002-7863

DOI - 10.1021/ja902681k

Subject(s) - chemistry , sortase a , sortase , biochemistry , terminal (telecommunication) , stereochemistry , bacterial protein , gene , telecommunications , computer science

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.

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