Site-Specific Protein Modification on Living Cells Catalyzed by Sortase

The use of enzymes is a promising approach for site-specific protein modification on living cells owing to their substrate specificity. Herein we describe a general strategy for the site-specific modification of cell surface proteins with synthetic molecules by using Sortase, a transpeptidase from S...

Full description

Saved in:
Bibliographic Details
Published in:Chembiochem : a European journal of chemical biology 2008-03, Vol.9 (5), p.802-807
Main Authors: Tanaka, Tsutomu, Yamamoto, Teruyasu, Tsukiji, Shinya, Nagamune, Teruyuki
Format: Article
Language:English
Subjects:
Aminoacyltransferases - chemistry
Bacterial Proteins - chemistry
Biotin - chemistry
Catalysis
cell surface engineering
Cell Survival
Cells, Cultured
Cysteine Endopeptidases - chemistry
Fluorescent Dyes - chemistry
Humans
Membrane Proteins - chemistry
Molecular Structure
peptide tags
Peptides - chemistry
Protein Binding
protein-protein conjugation
RANK Ligand - chemistry
site-specific labeling
Sortase
Staphylococcus aureus
Staphylococcus aureus - enzymology
Substrate Specificity
Surface Properties
Time Factors
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The use of enzymes is a promising approach for site-specific protein modification on living cells owing to their substrate specificity. Herein we describe a general strategy for the site-specific modification of cell surface proteins with synthetic molecules by using Sortase, a transpeptidase from Staphylococcus aureus. The short peptide tag LPETGG is genetically introduced to the C terminus of the target protein, expressed on the cell surface. Subsequent addition of Sortase and an N-terminal triglycine-containing probe results in the site-specific labeling of the tagged protein. We were successful in the C-terminal-specific labeling of osteoclast differentiation factor (ODF) with a biotin- or fluorophore-containing short peptide on the living cell surface. The labeling reaction occurred efficiently in serum-containing medium, as well as serum-free medium or PBS. The labeled products were detected after incubation for 5 min. In addition, site-specific protein-protein conjugation was successfully demonstrated on a living cell surface by the Sortase-catalyzed reaction. This strategy provides a powerful tool for cell biology and cell surface engineering.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.200700614