Binding specificity of the lantibiotic-binding immunity protein NukH

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

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Abstract

NukH is a lantibiotic-binding immunity protein that shows strong binding activity against type A(II) lantibiotics. In this study, the binding specificity of NukH was analyzed by using derivatives of nukacin ISK-1, which is a type A(II) lantibiotic produced by Staphylococcus warneri ISK-1. Interactions between cells of Lactococcus lactis transformants expressing nukH and nukacin ISK-1 derivatives were analyzed by using a quantitative peptide-binding assay. Differences in the cell-binding rates of each nukacin ISK-1 derivative suggested that three lysine residues at positions 1 to 3 of nukacin ISK-1 contribute to the effective binding of nukacin ISK-1 to nukH-expressing cells. The binding levels of mutants with lanthionine and dehydrobutyrine substitutions (S11A nukacin4-27 and T24A nukacin4-27, respectively) to nukH-expressing cells were considerably lower than those of nukacin 4-27, suggesting that unusual amino acids play a decisive role in NukH recognition. Additionally, it was suggested that T9A nukacin 4-27, a mutant with a 3-methyllanthionine substitution, binds to NukH via an intermolecular disulfide bond after it is weakly recognized by NukH. We succeeded in the detection of specific type A(II) lantibiotics from the culture supernatants of various bacteriocin producers by using the binding specificity of nukH-expressing cells.

Original languageEnglish
Pages (from-to)7613-7619
Number of pages7
JournalApplied and environmental microbiology
Volume74
Issue number24
DOIs
Publication statusPublished - Dec 2008

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

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    Okuda, K. I., Yanagihara, S., Shioya, K., Harada, Y., Nagao, J. I., Aso, Y., Zendo, T., Nakayama, J., & Sonomoto, K. (2008). Binding specificity of the lantibiotic-binding immunity protein NukH. Applied and environmental microbiology, 74(24), 7613-7619. https://doi.org/10.1128/AEM.00789-08