Microidentification of N-terminal-blocked amino acid residues of proteins and peptides

Yasuhiko Kawakami, Shinji Ohmori

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Proteins or peptides having an N-terminal-blocked amino acid were successively digested by pronase E, proteinase K, and carboxypeptidase Y. The N-blocked amino acids released from proteins or peptides were derivatized with 9-anthryldiazomethane (ADAM) to the corresponding esters followed by addition of formic acid to remove the remaining ADAM which interfered with further analysis. The anthryl esters were analyzed by high-performance liquid chromatography equipped with a fluorimetric detector. Twelve N-acetylamino acids (Asn, Gln, Ser, Thr, Gly, Ala, Tyr, Pro, Met, Val, Ile, and Leu), pyroglutamic acid, N-formylMet, and N-myristoylGly could be separated from each other and identified on the same chromatographic run. As examples of applied experiments to proteins and peptides, N-acetyl derivatives of Ser, Ala, Met, Gly, Tyr, and Pro as well as N-myristoylGly could be satisfactorily identified using 100 pmol each of seven proteins and peptides. The method reported here is an improved one that was reported in the previous paper based on the same principles.

Original languageEnglish (US)
Pages (from-to)66-72
Number of pages7
JournalAnalytical Biochemistry
Volume220
Issue number1
DOIs
StatePublished - Jan 1 1994
Externally publishedYes

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