Functional significance of the E loop, a novel motif conserved in the lantibiotic immunity ATP-binding cassette transport systems.
Lantibiotics are peptide-derived antibacterial substances produced by some Gram-positive bacteria and characterized by the presence of unusual amino acids, like lanthionines and dehydrated amino acids. Because lantibiotic producers may be attacked by self-produced lantibiotics, they express immunity proteins on the cytoplasmic membrane. An ATP-binding cassette (ABC) transport system mediated by the LanFEG protein complex is a major system in lantibiotic immunity. Multiple-sequence alignment analysis revealed that LanF proteins contain the E loop, a variant of the Q loop, which is a well-conserved motif in the nucleotide-binding domains (NBDs) of general ABC transporters. To elucidate E loop function, we introduced a mutation in the NukF protein, which is involved in the nukacin-ISK-1 immunity system. Amino acid replacement of glutamic acid in the E loop with glutamine (E85Q) resulted in slight decreases in the immunity level and transport activity. Additionally, the E85A mutation severely impaired the immunity level and transport activity. On the other hand, ATPase activities of purified E85Q and E85A mutants were almost similar to that of the wild type. These results suggested that the E loop found in ABC transporters involved in lantibiotic immunity plays a significant role in the function of these transporters, especially in the structural change of transmembrane domains.
Okuda
Ken-ichi
K
Department of Bioscience & Biotechnology, Faculty of Agriculture, Graduate School, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
Yanagihara
Sae
S
Sugayama
Tomomichi
T
Zendo
Takeshi
T
Nakayama
Jiro
J
Sonomoto
Kenji
K
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D020816:Amino Acid Motifs / Q000235:genetics / Q000502:physiology*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001430:Bacteriocins / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects / Q000235:genetics; D015153:Blotting, Western; D024881:Drug Resistance, Bacterial / Q000235:genetics / Q000502:physiology; D007778:Lactobacillus / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D013294:Lactococcus lactis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D001430:Bacteriocins; C421186:nukacin ISK-1
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