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A Minimalist Substrate for Enzymatic Peptide and Protein Conjugation
Author(s) -
Wollack James W.,
Silverman Julie M.,
Petzold Christopher J.,
Mougous Joseph D.,
Distefano Mark D.
Publication year - 2009
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200900566
Subject(s) - prenylation , farnesyltransferase , cysteine , peptide , chemistry , amino acid , residue (chemistry) , combinatorial chemistry , stereochemistry , substrate (aquarium) , enzyme , biochemistry , biology , ecology
Recently a number of nonnatural prenyl groups containing alkynes and azides have been developed as handles to perform click chemistry on proteins and peptides ending in the sequence “CAAX”, where C is a cysteine that becomes alkylated, A is an aliphatic amino acid and X is any amino acid. When such molecules are modified, a tag containing a prenyl analogue and the “CAAX box” sequence remains. Here we report the synthesis of an alkyne‐containing substrate comprised of only nine nonhydrogen atoms. This substrate was synthesized in six steps from 3‐methylbut‐2‐en‐1‐ol and has been enzymatically incorporated into both proteins and peptides by using protein farnesyltransferase. After prenylation the final three amino acids required for enzymatic recognition can be removed by using carboxypeptidase Y, leaving a single residue (the cysteine from the “CAAX box”) and the prenyl analogue as the only modifications. We also demonstrate that this small tag minimizes the impact of the modification on the solubility of the targeted protein. Hence, this new approach should be useful for applications in which the presence of a large tag hinders the modified protein’s solubility, reactivity, or utility.