Protein semi-synthesis in living cells.
Incorporation of chemical probes into proteins is a powerful way to elucidate biological processes and to engineer novel function. Here we describe an approach that allows ligation of synthetic molecules to target proteins in an intracellular environment. A cellular protein is genetically tagged with one-half of a split intein. The complementary half is linked in vitro to the synthetic probe, and this fusion is delivered into cells using a transduction peptide. Association of the intein halves in the cytosol triggers protein trans-splicing, resulting in the ligation of the probe to the target protein through a peptide bond. This process is specific and applicable to cytosolic and integral membrane proteins. The technology should allow cellular proteins to be elaborated with a variety of abiotic probes.
Giriat
Izabela
I
Laboratory of Synthetic Protein Chemistry, The Rockefeller University, New York, New York 10021, USA.
Muir
Tom W
TW
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004258:DNA Polymerase III / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D014176:Protein Biosynthesis; D019154:Protein Splicing; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D014162:Transfection
D008164:Luminescent Proteins; D011506:Proteins; D049452:Green Fluorescent Proteins; C074904:DNA polymerase III, alpha subunit; D004258:DNA Polymerase III