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 |
2008-12-1
|
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 |
4.016
|
Times Cited |
10
|
WOS Category
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
MICROBIOLOGY
|
Resource |
General Microbes |
JCM1157 |