A method for the detection of asparagine deamidation and aspartate isomerization of proteins by MALDI/TOF-mass spectrometry using endoproteinase Asp-N

Daisuke Kameoka, Tadashi Ueda, Taiji Imoto

研究成果: Contribution to journalArticle査読

29 被引用数 (Scopus)

抄録

A method was established for evaluating Asn deamidation and Asp isomerization/racemization. To detect the subtle changes in mass that accompany these chemical modifications, we used a combination of enzyme digestion by endoproteinase Asp-N, which selectively cleaves the N-terminus of L-α-Asp, and MALDI/TOF-mass spectrometry. To achieve better resolution, we employed digests of 15N-labeled protein as an internal standard. To demonstrate the advantages of this method, we applied it to identify deamidated sites in mutant lysozymes in which the Asn residue is mutated to Asp. We also identified the deamidation or isomerization site of the lysozyme samples after incubating them under acidic or basic conditions.

本文言語英語
ページ(範囲)129-135
ページ数7
ジャーナルJournal of Biochemistry
134
1
DOI
出版ステータス出版済み - 7 1 2003

All Science Journal Classification (ASJC) codes

  • Medicine(all)
  • Biochemistry
  • Molecular Biology

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