title: The presence of modifiable residues in the core peptide part of precursor nisin is not crucial for precursor nisin interactions with NisB- and NisC.
abstract: Precursor nisin is a model posttranslationally modified precursor lantibiotic that can be structurally divided into a leader peptide sequence and a modifiable core peptide part. The nisin core peptide clearly plays an important role in the precursor nisin-nisin modification enzymes interactions, since it has previously been shown that the construct containing only the nisin leader sequence is not sufficient to pull-down the nisin modification enzymes NisB and NisC. Serines and threonines in the core peptide part are the residues that NisB specifically dehydrates, and cysteines are the residues that NisC stereospecifically couples to the dehydrated amino acids. Here, we demonstrate that increasing the number of negatively charged residues in the core peptide part of precursor nisin, which are absent in wild-type nisin, does not abolish binding of precursor nisin to the modification enzymes NisB and NisC, but dramatically decreases the antimicrobial potency of these nisin mutants. An unnatural precursor nisin variant lacking all serines and threonines in the core peptide part and an unnatural precursor nisin variant lacking all cysteines in the core peptide part still bind the nisin modification enzymes NisB and NisC, suggesting that these residues are not essential for direct interactions with the nisin modification enzymes NisB and NisC. These results are important for lantibiotic engineering studies.
authors.affiliation: Molecular Genetics Department, University of Groningen, Groningen, The Netherlands ; Synthetic Biology Centre, University of Groningen, Groningen, The Netherlands.
authors.forename: Rustem
authors.initials: R
authors.lastname: Khusainov
authors.forename: Oscar P
authors.initials: OP
authors.lastname: Kuipers
mesh_terms: D000890:Anti-Infective Agents / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D003545:Cysteine / Q000737:chemistry; D014644:Genetic Variation; D019753:Intramolecular Lyases / Q000737:chemistry* / Q000235:genetics; D013294:Lactococcus lactis / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics; D009561:Nisin / Q000737:chemistry* / Q000235:genetics; D010455:Peptides / Q000737:chemistry; D010957:Plasmids; D011485:Protein Binding; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011995:Recombination, Genetic; D012694:Serine / Q000737:chemistry; D013379:Substrate Specificity; D013912:Threonine / Q000737:chemistry
chemical_list: D000890:Anti-Infective Agents; D001426:Bacterial Proteins; D008565:Membrane Proteins; C079126:NisB protein, Lactococcus lactis; D010455:Peptides; D009561:Nisin; D013912:Threonine; D012694:Serine; D019753:Intramolecular Lyases; D003545:Cysteine
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