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C-terminal Sequence Analysis of Proteins by MALDI TOF MS Using Selective C-terminal Derivatization with Hydrazine and Bromine-containing Nucleophile

등록일
2008년 8월 12일 09시 15분 38초
접수번호
0981
발표코드
32P83포 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
발표시간
금 <발표Ⅳ>
발표형식
포스터
발표분야
분석화학
저자 및
공동저자
안성희, 김종서, 김희준
서울대학교 화학부, Korea
C-terminal sequence analysis of proteins and peptides serves as a powerful tool for understanding the interaction between proteins. Unfortunately, there are only few methods for determining the C-terminal sequence, which are mostly limited to large scale experiments. This is because the carboxyl group is difficult to modify, so the yield of C-terminal derivatized peptides is very low. Here, we introduce a new nanomole scale method that easily modifies the C-terminus by activating it into an active ester through an oxazolone ring in a mixture of acetic anhydride and formic acid. It is then derivatized with a strong nucleophile like bromophenylhydrazine. The isotopic signature from the bromine atom is useful for visual recognition of the y-series of the bromine-containing C-terminus. MALDI-TOF MS analysis of several model peptides and proteins, derivatized by the described method, yielded intense m/z peaks of the expected peptide ions. MSMS analysis is also in progress.

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