Protein Science, Vol 7, Issue 8 1839-1842, Copyright © 1998 by Cold Spring Harbor Laboratory Press

FOR THE RECORD

Overexpression of recombinant proteins with a C-terminal thiocarboxylate: Implications for protein semisynthesis and thiamin biosynthesis

C. KINSLAND, S. V. TAYLOR, N. L. KELLEHER, F. W. MCLAFFERTY and T. P. BEGLEY
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853

A facile and rapid method for the production of protein C-terminal thiocarboxylates on DNA-encoded polypeptides is described. This method, which relies on the mechanism of the cleavage reaction of intein-containing fusion proteins, can produce multimilligram quantities of protein C-terminal thiocarboxylate quickly and inexpensively. The utility of this method for protein semisynthesis and implications for studies on the biosynthesis of thiamin are discussed.
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