Localization and interaction of the biosynthetic proteins for the lantibiotic, Nukacin ISK-1.
Nukacin ISK-1 is a type-A(II) lantibiotic produced by Staphylococcus warneri ISK-1. In this study, we characterized NukM and NukT, which are predicted to be involved in modification of prepeptide (NukA) and cleavage of leader peptide and subsequent secretion respectively. Localization analysis of NukM and NukT in the wild-type strain indicated that both proteins were located at the cytoplasm membrane. Interestingly, NukM expressed heterologously in St. carnosus TM300 was also located at the cytoplasm membrane even in the absence of NukT. Yeast two-hybrid assay showed that a complex of at least two each of NukM and NukT was associated with NukA. In vitro interaction analysis by surface plasmon resonance biosensor further suggested that membrane-located NukM interacted with NukA. These results indicate that NukM and NukT form a membrane-located multimeric protein complex and that post-translational modification of nukacin ISK-1 would occur at the cytoplasm membrane.
Nagao
Jun-ichi
J
Laboratory of Microbial Technology, Division of Microbial Science and Technology, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University Graduate School, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
Aso
Yuji
Y
Sashihara
Toshihiro
T
Shioya
Kouki
K
Adachi
Asaho
A
Nakayama
Jiro
J
Sonomoto
Kenji
K
D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001430:Bacteriocins / Q000096:biosynthesis*; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013210:Staphylococcus / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins; D001430:Bacteriocins; C421186:nukacin ISK-1