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C‐terminal labelling of recombinant proteins using an engineered split‐intein
Author(s) -
Volkmann Gerrit,
Liu XiangQin
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.863.7
Subject(s) - intein , protein tag , labelling , protein splicing , rna splicing , recombinant dna , peptide , protein engineering , biochemistry , chemistry , biotin , computational biology , biology , fusion protein , gene , enzyme , rna
Site‐specific labelling of proteins has great potential to aid in the characterization of protein function, three‐dimensional structure, folding behaviour, and interaction with other proteins or small‐molecules. However, new and more efficient labelling methods are needed to satisfy the increasing demand for creating site‐specifically modified proteins. Here, a novel technique is presented to specifically label recombinant proteins at the C‐terminus using a peptide‐protein trans ‐splicing approach. A label is incorporated into a small synthetic peptide, and then transferred onto the C‐terminus of the protein of interest via a stable peptide bond formed by protein trans ‐splicing. We applied this method to introduce fluorescent labels and biotin at the C‐terminus of recombinant proteins, with biotin being potentially useful in creating protein microchips. The gentle chemistry of the trans ‐splicing method also allowed us to label receptor proteins on the surface of eukaryotic cells, indicating a broad applicability of our labelling technique. Research support was provided by CIHR and NSERC.