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: 著書/レポートタイプへの貢献

9 引用 (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.

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

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Bacteriocins
Ligases
Amino Acids
Peptides
Histidine
Gram-Positive Bacteria
Enzymes
Affinity Chromatography
Mass Spectrometry
Escherichia coli
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics
  • Medicine(all)

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

Engineering unusual amino acids into peptides using lantibiotic synthetase. / Nagao, Jun Ichi; Shioya, Kouki; Harada, Yoshitaka; Okuda, Ken Ichi; Zendo, Takeshi; Nakayama, Jiro; Sonomoto, Kenji.

Heterologous Gene Expression in E.coli: Methods and Protocols. 版 / Thomas Evans, Jr.; Ming-Qun Xu. 2011. p. 225-236 (Methods in Molecular Biology; 巻 705).

研究成果: 著書/レポートタイプへの貢献

Nagao, JI, Shioya, K, Harada, Y, Okuda, KI, 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. Methods in Molecular Biology, 巻. 705, pp. 225-236. https://doi.org/10.1007/978-1-61737-967-3_13
Nagao JI, Shioya K, Harada Y, Okuda KI, Zendo T, Nakayama J その他. Engineering unusual amino acids into peptides using lantibiotic synthetase. : Evans, Jr. T, Xu M-Q, 編集者, Heterologous Gene Expression in E.coli: Methods and Protocols. 2011. p. 225-236. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61737-967-3_13
Nagao, Jun Ichi ; Shioya, Kouki ; Harada, Yoshitaka ; Okuda, Ken Ichi ; Zendo, Takeshi ; Nakayama, Jiro ; Sonomoto, Kenji. / Engineering unusual amino acids into peptides using lantibiotic synthetase. Heterologous Gene Expression in E.coli: Methods and Protocols. 編集者 / Thomas Evans, Jr. ; Ming-Qun Xu. 2011. pp. 225-236 (Methods in Molecular Biology).
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