LiaRS reporter assay: A simple tool to identify lipid II binding moieties in lantibiotic nukacin ISK-1

Khaled M Elsayed; Mohammad R Islam; Abdullah-Al-Mahin; Jun-Ichi Nagao; Takeshi Zendo; Kenji Sonomoto

doi:10.1016/j.jbiosc.2016.10.002

LiaRS reporter assay: A simple tool to identify lipid II binding moieties in lantibiotic nukacin ISK-1

J Biosci Bioeng. 2017 Mar;123(3):398-401. doi: 10.1016/j.jbiosc.2016.10.002. Epub 2016 Nov 14.

Authors

Khaled M Elsayed¹, Mohammad R Islam², Abdullah-Al-Mahin³, Jun-Ichi Nagao⁴, Takeshi Zendo¹, Kenji Sonomoto⁵

Affiliations

¹ Laboratory of Microbial Technology, Division of Systems Bioengineering, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
² Laboratory of Microbial Technology, Division of Systems Bioengineering, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan; Department of Biochemistry and Molecular Biology, Faculty of Biological Sciences, University of Dhaka, Dhaka 1000, Bangladesh.
³ Laboratory of Microbial Technology, Division of Systems Bioengineering, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan; Microbiology and Industrial Irradiation Division, Institute of Food and Radiation Biology, Atomic Energy Research Establishment, Savar, Dhaka 1349, Bangladesh.
⁴ Section of Infection Biology, Department of Functional Bioscience, Fukuoka Dental College, 2-15-1 Tamura, Sawara-ku, Fukuoka 814-0913, Japan.
⁵ Laboratory of Microbial Technology, Division of Systems Bioengineering, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan. Electronic address: [email protected].

PMID: 27856233
DOI: 10.1016/j.jbiosc.2016.10.002

Abstract

Binding to lipid II is an important step in the mode of action of most lantibiotics targeting the bacterial cell wall. We applied the Bacillus subtilis two-component system, LiaRS, that is known to respond to antibiotics interfering with lipid II cycle, in order to evaluate lipid II binding activity of known bacteriocins and also to identify lipid II binding moieties in lantibiotic nukacin ISK-1. Using this method, we confirmed that the methyllanthionine ring in nukacin ISK-1 is crucial for lipid II binding as previously indicated. In this study, we further identified that the three N-terminal lysine residues (K1, K2, and K3) and the glycine (G5) residue in nukacin ISK-1 are also important in lipid II binding.

Keywords: Bacteriocin; Lantibiotic; Lipid II; Nukacin ISK-1; Structure-activity relationship; Two-component system.

MeSH terms

Alanine / analogs & derivatives
Alanine / metabolism
Amino Acid Sequence
Bacillus subtilis / metabolism*
Bacteriocins / chemistry*
Bacteriocins / metabolism*
Cell Wall / metabolism
Genes, Reporter*
Sulfides / metabolism
Uridine Diphosphate N-Acetylmuramic Acid / analogs & derivatives*
Uridine Diphosphate N-Acetylmuramic Acid / metabolism

Substances

Bacteriocins
Sulfides
Uridine Diphosphate N-Acetylmuramic Acid
muramyl-NAc-(pentapeptide)pyrophosphoryl-undecaprenol
nukacin ISK-1
beta-methyllanthionine
Alanine