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A Method for Identifying the Carboxy Terminal Amino Acid of a Protein

✍ Scribed by T. Hayashi; T. Sasagawa

Publisher: Elsevier Science
Year: 1993
Tongue: English
Weight: 437 KB
Volume: 209
Category: Article
ISSN: 0003-2697
DOI: 10.1006/abio.1993.1097

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✦ Synopsis

A highly sensitive new method for identifying the carboxy terminus of a protein was developed. The carboxyl terminal amino acid was racemized by reaction with acetic anhydride. The resulting modified protein was subjected to acid hydrolysis. The hydrolysate was derivatized with (+)-1-(9-fluorenyl)ethyl chloroformate to give fluorescent amino acid diastereomers. The amino acid diastereomers were separated on a reversed-phase column. Only carboxyl terminal amino acids give a D-amino acid. Application of this method was described for the isolation and identification of carboxyl terminal peptides from an enzymatic digest of a protein.

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