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

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• 作者：John M. Antos ; Guo-Liang Chew ; Carla P. Guimaraes ; Nicholas C. Yoder ; Gijsbert M. Grotenbreg ; Maximilian Wei-Lin Popp ; Hidde L. Ploegh
• 刊名：Journal of the American Chemical Society
• 出版年：2009
• 出版时间：August 12, 2009
• 年：2009
• 卷：131
• 期：31
• 页码：10800-10801
• 全文大小：152K
• 年卷期：v.131,no.31(August 12, 2009)
• ISSN：1520-5126

文摘

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.