The Perkin-elmer Corporation

Applied Biosystems, LLC

MeriLisa Bozzini

Victoria L Boyd

Gerald Zon

Robert J DeFranco

G Marc Loudon

Piotr J Guga

A method of forming a thiohydantoin from an N-protected amino acid is described. The method employs a phosphate compound selected from the group consisting of

Thiohydantoin formation and selective modification of the carboxy

A method is provided for C-terminal sequencing of a protein or peptide. An important feature of the method is the formation of an oxazolone moiety at the C-terminus of a protein or peptide by treatment with acetic anhydride under basic conditions followed by conversion of the oxazolone to a thiohydantoin moiety by treatment with thiocyanate under acidic conditions. Yields of thiohydantoin are further enhanced by delivering thiocyanate as the conjugate acid of a sterically hindered alkylammnonium cation.

Acetic anhydride activation for C-terminal protein sequencing

A method of forming a thiohydantoin from an N-protected amino acid. The method employs a uronium or phosphonium compound to activate the terminal carboxyl group of the amino acid and a thiocyanate reagent to cyclize the activated amino acid to the thio-hydantoin. The thiohydantoin can be cleaved from its N-protecting group, for use in C-terminal peptide sequencing. Particularly preferred uronium compounds include salts of 2-chlorouronium. Preferred thiocyanate reagents include trimethylsilyl isothiocyanate and crown ether adducts of metallothiocyanates, such as the 18-crown-6 adduct of KSCN.

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Burlingame, CA, United States of America

MeriLisa Bozzini