Enzymatic incorporation of orthogonally reactive prenylazide groups into peptides using geranylazide diphosphate via protein farnesyltransferase: implications for selective protein labeling.
Protein farnesyltransferase (PFTase) catalyzes the attachment of a geranyl azide moiety to a peptide substrate, N-dansyl-Gly-Cys-Val-Ile-Ala-OH. The resulting azide-containing peptide was derivatized with a triphenylphosphine-based reagent to generate an O-alkyl imidate-linked product, rather than the amide-linked material expected via a Staudinger reaction. Since the CAAX box recognition motif (where the internal A residues are aliphatic amino acids) modified by PFTase can be incorporated into the C-terminus of virtually any polypeptide, this two-step procedure provides a general method for incorporating a diverse range of chemical modifications specifically near the C-terminus of proteins.
Rose
Matthew W
MW
Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA.
Xu
Juhua
J
Kale
Tamara A
TA
O'Doherty
George
G
Barany
George
G
Distefano
Mark D
MD
D019883:Alkyl and Aryl Transferases / Q000378:metabolism*; D001386:Azides / Q000737:chemistry; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D011106:Polyisoprenyl Phosphates / Q000737:chemistry; D017368:Protein Prenylation; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*
D001386:Azides; D007202:Indicators and Reagents; D011106:Polyisoprenyl Phosphates; D011506:Proteins; D019883:Alkyl and Aryl Transferases; C064911:p21(ras) farnesyl-protein transferase