RRC ID 5160
Author Okuda K, Yanagihara S, Shioya K, Harada Y, Nagao J, Aso Y, Zendo T, Nakayama J, Sonomoto K.
Title Binding specificity of the lantibiotic-binding immunity protein NukH.
Journal Appl. Environ. Microbiol.
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 nukacin(4-27) and T24A nukacin(4-27), 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.
Volume 74(24)
Pages 7613-9
Published 2008-12
DOI 10.1128/AEM.00789-08
PII AEM.00789-08
PMID 18978082
PMC PMC2607159
MeSH Bacterial Proteins / genetics Bacterial Proteins / metabolism* Bacteriocins / genetics Bacteriocins / metabolism* Lactococcus lactis / genetics Lactococcus lactis / metabolism* Models, Molecular Mutagenesis, Site-Directed Mutant Proteins / genetics Mutant Proteins / metabolism Mutation, Missense Protein Binding Protein Conformation
IF 3.633
Times Cited 7
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY
Resource
General Microbes JCM1157