Influence of amino acid substitutions in the nisin leader peptide on biosynthesis and secretion of nisin by Lactococcus lactis.

Détails

ID Serval
serval:BIB_EC50502BCA09
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Influence of amino acid substitutions in the nisin leader peptide on biosynthesis and secretion of nisin by Lactococcus lactis.
Périodique
Journal of Biological Chemistry
Auteur(s)
van der Meer J.R., Rollema H.S., Siezen R.J., Beerthuyzen M.M., Kuipers O.P., de Vos W.M.
ISSN
0021-9258 (Print)
ISSN-L
0021-9258
Statut éditorial
Publié
Date de publication
1994
Peer-reviewed
Oui
Volume
269
Numéro
5
Pages
3555-3562
Langue
anglais
Résumé
Structural genes for small lanthionine-containing antimicrobial peptides, known as lantibiotics, encode N-terminal leader sequences which are not present in the mature peptide, but are cleaved off at some stage in the maturation process. Leader sequences of the different lantibiotics share a number of identical amino acid residues, but they are clearly different from sec-dependent protein export signal sequences. We studied the role of the leader sequence of the lantibiotic nisin, which is produced and secreted by Lactococcus lactis, by creating site-directed mutations at various positions in the leader peptide sequence. Mutations at Arg-1 and Ala-4, but not at the conserved Pro-2, strongly affected the processing of the leader sequence and resulted in the extracellular accumulation of a biologically inactive precursor peptide. Amino acid analysis and 1H NMR studies indicated that the precursor peptide with an Ala-4-->Asp mutation contained a modified nisin structural part with the (mutated) unmodified leader sequence still attached to it. The Ala-4-->Asp precursor peptide could be activated in vitro by enzymatic cleavage with trypsin, liberating nisin. These results confirmed that cleavage of the leader peptide is the last step in nisin maturation and is necessary to generate a biologically active peptide. Several mutations, i.e. Pro-2-->Gly,Pro-2-->Val, Asp-7-->Ala,Lys-9-->Leu,Ser-10-->Ala/Ser-12-->Ala and Val-11-->Asp/Val-13-->Glu in the leader peptide did not have any detectable effect on nisin production and secretion, although some of them affected highly conserved residues. When mutations were created in the -18 to -15 region of the nisin leader peptide (i.e. Phe-18-->Leu,Leu-16-->Lys,Asp-15-->Ala), no secretion or intracellular accumulation could be detected of nisin or its precursors. This suggested that these conserved residues are involved in the maturation process and may interact with lantibiotic-specific modifying enzymes.
Mots-clé
Amino Acid Sequence, Base Sequence, Chromatography, High Pressure Liquid, DNA Primers, Genes, Bacterial, Lactococcus lactis/metabolism, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Mutagenesis, Site-Directed, Nisin/biosynthesis, Nisin/chemistry, Plasmids, Point Mutation, Protein Biosynthesis, Protein Precursors/biosynthesis, Protein Precursors/chemistry, Protein Sorting Signals/genetics, Protein Sorting Signals/metabolism, Protein Structure, Secondary
Pubmed
Web of science
Création de la notice
21/01/2008 13:36
Dernière modification de la notice
20/08/2019 16:14
Données d'usage