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 引用 (Scopus)

抜粋

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.

元の言語英語
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
出版物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(印刷物)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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  • これを引用

    Nagao, J. I., Shioya, K., Harada, Y., Okuda, K. I., Zendo, T., Nakayama, J., & Sonomoto, K. (2011). Engineering unusual amino acids into peptides using lantibiotic synthetase. : T. Evans, Jr., & M-Q. Xu (版), Heterologous Gene Expression in E.coli: Methods and Protocols (pp. 225-236). (Methods in Molecular Biology; 巻数 705). https://doi.org/10.1007/978-1-61737-967-3_13