Engineering unusual amino acids into peptides using lantibiotic synthetase

Jun Ichi Nagao, Kouki Shioya, Yoshitaka Harada, Ken Ichi Okuda, Takeshi Zendo, Jiro Nakayama, Kenji Sonomoto

研究成果: Chapter in Book/Report/Conference proceedingChapter

9 被引用数 (Scopus)

抄録

Alteration of protein structure and function by introducing unusual amino acids has great potential to develop new biological tool and to produce novel therapeutic agents. Lantibiotics produced by Gram-positive bacteria are ribosomally synthesized and post-translationally modified antimicrobial peptides. The modification enzyme involved in lantibiotic biosynthesis can catalyze the formation of unusual amino acids in the nascent lantibiotic prepeptide. Here, a novel methodology on the lantibiotic nukacin ISK-1 is described for engineering unusual amino acid residues into hexa-histidine-tagged (His-tagged) prepeptide NukA by the modification enzyme NukM in Escherichia coli. Co-expression of His-tagged NukA and NukM, purification of the resulting His-tagged prepeptide by affinity chromatography, and subsequent mass spectrometry analysis show that the prepeptide is converted into a postulated peptide with decrease in mass which results from the formation of unusual amino acids such as dehydrated amino acid, lanthionine, or 3-methyl lanthionine at the expected positions. The modified prepeptide can be readily obtained by one-step purification. This strategy will thus be a simple and powerful tool for introducing unusual amino acid residues aimed at peptide engineering.

本文言語英語
ホスト出版物のタイトルHeterologous Gene Expression in E.coli
ホスト出版物のサブタイトルMethods and Protocols
編集者Thomas Evans, Jr., Ming-Qun Xu
ページ225-236
ページ数12
DOI
出版ステータス出版済み - 12 1 2011

出版物シリーズ

名前Methods in Molecular Biology
705
ISSN(印刷版)1064-3745

All Science Journal Classification (ASJC) codes

  • 分子生物学
  • 遺伝学

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