1887

Abstract

The importance of 3D structuring in the N- and C-terminal ends of the two peptides (39-mer LcnG- and 35-mer LcnG-) that constitute the two-peptide bacteriocin lactococcin G was analysed by replacing residues in the end regions with the corresponding -isomeric residues. When assayed for antibacterial activity in combination with the complementary wild-type peptide, LcnG- with four -residues in its C-terminal region and LcnG- with four -residues in either its N- or its C-terminal region were relatively active (two- to 20-fold reduction in activity). 3D structuring of the C-terminal region in LcnG- and the C- and N-terminal regions in LcnG- is thus not particularly critical for retaining antibacterial activity, indicating that the 3D structure of these regions is not vital for interpeptide interactions or for interactions between the peptides and cellular components. The 3D structure of the N-terminal region in LcnG- may be more important, as LcnG- with four N-terminal -residues was the least active of these four peptides (10- to 100-fold reduction in activity). The results are consistent with a proposed structural model of lactococcin G in which LcnG- and - form a transmembrane parallel helix–helix structure involving approximately 20 residues in each peptide, starting near the N terminus of LcnG- and at about residue 13 in LcnG-. Upon expressing the lactococcin G immunity protein, sensitive target cells became resistant to all of these -residue-containing peptides. The end regions of the two lactococcin G peptides are consequently not involved in essential structure-dependent interactions with the immunity protein. The relatively high activity of most of the -residue-containing peptides suggests that bacteriocins with increased resistance to exopeptidases may be generated by replacing their N- and C-terminal residues with -residues.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.038430-0
2010-06-01
2020-08-03
Loading full text...

Full text loading...

/deliver/fulltext/micro/156/6/1883.html?itemId=/content/journal/micro/10.1099/mic.0.038430-0&mimeType=html&fmt=ahah

References

  1. Axelsson L., Holck A.. 1995; The genes involved in production of and immunity to sakacin A, a bacteriocin from Lactobacillus sake Lb706. J Bacteriol177:2125–2137
    [Google Scholar]
  2. Axelsson L., Katla T., Bjørnslett M., Eijsink V. G., Holck A.. 1998; A system for heterologous expression of bacteriocins in Lactobacillus sake. FEMS Microbiol Lett168:137–143
    [Google Scholar]
  3. Diep D. B., Skaugen M., Salehian Z., Holo H., Nes I. F.. 2007; Common mechanisms of target cell recognition and immunity for class II bacteriocins. Proc Natl Acad Sci U S A104:2384–2389
    [Google Scholar]
  4. Donermeyer D. L., Allen P. M.. 1989; Binding to Ia protects an immunogenic peptide from proteolytic degradation. J Immunol142:1063–1068
    [Google Scholar]
  5. Hauge H. H., Nissen-Meyer J., Nes I. F., Eijsink V. G.. 1998; Amphiphilic α-helices are important structural motifs in the α and β peptides that constitute the bacteriocin lactococcin G – enhancement of helix formation upon αβ interaction. Eur J Biochem251:565–572
    [Google Scholar]
  6. Håvarstein L. S., Diep D. B., Nes I. F.. 1995; A family of bacteriocin ABC transporters carry out proteolytic processing of their substrates concomitant with export. Mol Microbiol16:229–240
    [Google Scholar]
  7. Moll G., Ubbink-Kok T., Hildeng H. H., Nissen-Meyer J., Nes I. F., Konings W. N., Driessen A. J.. 1996; Lactococcin G is a potassium ion-conducting, two-component bacteriocin. J Bacteriol178:600–605
    [Google Scholar]
  8. Moll G., Hildeng H. H., Nissen-Meyer J., Nes I. F., Konings W. N., Driessen A. J.. 1998; Mechanistic properties of the two-component bacteriocin lactococcin G. J Bacteriol180:96–99
    [Google Scholar]
  9. Nes I. F., Håvarstein L. S., Holo H.. 1995; Genetics of non-lantibiotic bacteriocins. Dev Biol Stand85:645–651
    [Google Scholar]
  10. Nissen-Meyer J., Holo H., Håvarstein L. S., Sletten K., Nes I. F.. 1992; A novel lactococcal bacteriocin whose activity depends on the complementary action of two peptides. J Bacteriol174:5686–5692
    [Google Scholar]
  11. Nissen-Meyer J., Rogne P., Oppegård C., Haugen H. S., Kristiansen P. E.. 2009; Structure–function relationships of the non-lanthionine-containing peptide (class II) bacteriocins produced by Gram-positive bacteria. Curr Pharm Biotechnol10:19–37
    [Google Scholar]
  12. Oppegård C., Fimland G., Thorbek L., Nissen-Meyer J.. 2007; Analysis of the two-peptide bacteriocins lactococcin G and enterocin 1071 by site-directed mutagenesis. Appl Environ Microbiol73:2931–2938
    [Google Scholar]
  13. Oppegård C., Schmidt J., Kristiansen P. E., Nissen-Meyer J.. 2008; Mutational analysis of putative helix–helix interacting GxxxG-motifs and tryptophan residues in the two-peptide bacteriocin lactococcin G. Biochemistry47:5242–5249
    [Google Scholar]
  14. Oppegård C., Emanuelsen L., Thorbek L., Fimland G., Nissen-Meyer J.. 2010; The lactococcin G immunity protein recognizes specific regions in both peptides constituting the two-peptide bacteriocin lactococcin G. Appl Environ Microbiol76:1267–1273
    [Google Scholar]
  15. Powell M. F., Stewart T., Otvos L. Jr, Urge L., Gaeta F. C., Sette A., Arrhenius T., Thomson D., Soda K., Colon S. M.. 1993; Peptide stability in drug development. II. Effect of single amino acid substitution and glycosylation on peptide reactivity in human serum. Pharm Res10:1268–1273
    [Google Scholar]
  16. Rogne P., Fimland G., Nissen-Meyer J., Kristiansen P. E.. 2008; Three-dimensional structure of the two peptides that constitute the two-peptide bacteriocin lactococcin G. Biochim Biophys Acta 1784;543–554
    [Google Scholar]
  17. Tugyi R., Uray K., Ivan D., Fellinger E., Perkins A., Hudecz F.. 2005; Partial d-amino acid substitution: improved enzymatic stability and preserved Ab recognition of a MUC2 epitope peptide. Proc Natl Acad Sci U S A102:413–418
    [Google Scholar]
  18. Uteng M., Hauge H. H., Brondz I., Nissen-Meyer J., Fimland G.. 2002; Rapid two-step procedure for large-scale purification of pediocin-like bacteriocins and other cationic antimicrobial peptides from complex culture medium. Appl Environ Microbiol68:952–956
    [Google Scholar]
  19. van de Guchte M., van der Vossen J. M., Kok J., Venema G.. 1989; Construction of a lactococcal expression vector: expression of hen egg white lysozyme in Lactococcus lactissubsp. lactis. Appl Environ Microbiol55:224–228
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.038430-0
Loading
/content/journal/micro/10.1099/mic.0.038430-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error