Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Jan;179(1):135–140. doi: 10.1128/jb.179.1.135-140.1997

Role of transmembrane pH gradient and membrane binding in nisin pore formation.

G N Moll 1, J Clark 1, W C Chan 1, B W Bycroft 1, G C Roberts 1, W N Konings 1, A J Driessen 1
PMCID: PMC178671  PMID: 8981990

Abstract

Nisin is a cationic antimicrobial peptide that belongs to the group of lantibiotics. It is thought to form oligomeric pores in the target membrane by a mechanism that requires the transmembrane electrical potential delta psi and that involves local pertubation of the lipid bilayer structure. Here we show that nisin does not form exclusively voltage-dependent pores: even in the absence of a delta psi, nisin is able to dissipate the transmembrane pH gradient (delta pH) in sensitive Lactococcus lactis cells and proteoliposomes. The rate of dissipation increases with the magnitude of the delta pH. Nisin forms pores only when the delta pH is inside alkaline. The efficiency of delta psi-induced pore formation is strongly affected by the external pH, whereas delta pH-induced pore formation is rather insensitive to the external pH. Nisin(1-12), an amino-terminal fragment of nisin, and (des-deltaAla5)-(nisin(1-32) amide have a strongly reduced capacity to dissipate the delta psi and delta pH in cytochrome c oxidase proteoliposomes and L. lactis cells. Both variants bind with reduced efficiency to liposomes containing negatively charged phospholipids, suggesting that both ring A and rings C to E play a role in membrane binding. Nisin(1-12) competes with nisin for membrane binding and antagonizes pore formation. These findings are consistent with the wedge model of nisin-induced pore formation.

Full Text

The Full Text of this article is available as a PDF (223.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Batzri S., Korn E. D. Single bilayer liposomes prepared without sonication. Biochim Biophys Acta. 1973 Apr 16;298(4):1015–1019. doi: 10.1016/0005-2736(73)90408-2. [DOI] [PubMed] [Google Scholar]
  2. Bierbaum G., Sahl H. G. Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase. J Bacteriol. 1987 Dec;169(12):5452–5458. doi: 10.1128/jb.169.12.5452-5458.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bierbaum G., Sahl H. G. Induction of autolysis of staphylococci by the basic peptide antibiotics Pep 5 and nisin and their influence on the activity of autolytic enzymes. Arch Microbiol. 1985 Apr;141(3):249–254. doi: 10.1007/BF00408067. [DOI] [PubMed] [Google Scholar]
  4. Booth I. R. Regulation of cytoplasmic pH in bacteria. Microbiol Rev. 1985 Dec;49(4):359–378. doi: 10.1128/mr.49.4.359-378.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bruno M. E., Kaiser A., Montville T. J. Depletion of proton motive force by nisin in Listeria monocytogenes cells. Appl Environ Microbiol. 1992 Jul;58(7):2255–2259. doi: 10.1128/aem.58.7.2255-2259.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chan W. C., Dodd H. M., Horn N., Maclean K., Lian L. Y., Bycroft B. W., Gasson M. J., Roberts G. C. Structure-activity relationships in the peptide antibiotic nisin: role of dehydroalanine 5. Appl Environ Microbiol. 1996 Aug;62(8):2966–2969. doi: 10.1128/aem.62.8.2966-2969.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chan W. C., Leyland M., Clark J., Dodd H. M., Lian L. Y., Gasson M. J., Bycroft B. W., Roberts G. C. Structure-activity relationships in the peptide antibiotic nisin: antibacterial activity of fragments of nisin. FEBS Lett. 1996 Jul 22;390(2):129–132. doi: 10.1016/0014-5793(96)00638-2. [DOI] [PubMed] [Google Scholar]
  8. Demel R. A., Peelen T., Siezen R. J., De Kruijff B., Kuipers O. P. Nisin Z, mutant nisin Z and lacticin 481 interactions with anionic lipids correlate with antimicrobial activity. A monolayer study. Eur J Biochem. 1996 Jan 15;235(1-2):267–274. doi: 10.1111/j.1432-1033.1996.00267.x. [DOI] [PubMed] [Google Scholar]
  9. Dodd H. M., Horn N., Giffard C. J., Gasson M. J. A gene replacement strategy for engineering nisin. Microbiology. 1996 Jan;142(Pt 1):47–55. doi: 10.1099/13500872-142-1-47. [DOI] [PubMed] [Google Scholar]
  10. Driessen A. J., Konings W. N. Insertion of lipids and proteins into bacterial membranes by fusion with liposomes. Methods Enzymol. 1993;221:394–408. doi: 10.1016/0076-6879(93)21032-4. [DOI] [PubMed] [Google Scholar]
  11. Driessen A. J., van den Hooven H. W., Kuiper W., van de Kamp M., Sahl H. G., Konings R. N., Konings W. N. Mechanistic studies of lantibiotic-induced permeabilization of phospholipid vesicles. Biochemistry. 1995 Feb 7;34(5):1606–1614. doi: 10.1021/bi00005a017. [DOI] [PubMed] [Google Scholar]
  12. Gao F. H., Abee T., Konings W. N. Mechanism of action of the peptide antibiotic nisin in liposomes and cytochrome c oxidase-containing proteoliposomes. Appl Environ Microbiol. 1991 Aug;57(8):2164–2170. doi: 10.1128/aem.57.8.2164-2170.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Garcerá M. J., Elferink M. G., Driessen A. J., Konings W. N. In vitro pore-forming activity of the lantibiotic nisin. Role of protonmotive force and lipid composition. Eur J Biochem. 1993 Mar 1;212(2):417–422. doi: 10.1111/j.1432-1033.1993.tb17677.x. [DOI] [PubMed] [Google Scholar]
  14. Gross E., Morell J. L. The structure of nisin. J Am Chem Soc. 1971 Sep 8;93(18):4634–4635. doi: 10.1021/ja00747a073. [DOI] [PubMed] [Google Scholar]
  15. Kashket E. R. The proton motive force in bacteria: a critical assessment of methods. Annu Rev Microbiol. 1985;39:219–242. doi: 10.1146/annurev.mi.39.100185.001251. [DOI] [PubMed] [Google Scholar]
  16. Konings W. N., Poolman B., Driessen A. J. Bioenergetics and solute transport in lactococci. Crit Rev Microbiol. 1989;16(6):419–476. doi: 10.3109/10408418909104474. [DOI] [PubMed] [Google Scholar]
  17. Kuipers O. P., Beerthuyzen M. M., Siezen R. J., De Vos W. M. Characterization of the nisin gene cluster nisABTCIPR of Lactococcus lactis. Requirement of expression of the nisA and nisI genes for development of immunity. Eur J Biochem. 1993 Aug 15;216(1):281–291. doi: 10.1111/j.1432-1033.1993.tb18143.x. [DOI] [PubMed] [Google Scholar]
  18. Kuipers O. P., Beerthuyzen M. M., de Ruyter P. G., Luesink E. J., de Vos W. M. Autoregulation of nisin biosynthesis in Lactococcus lactis by signal transduction. J Biol Chem. 1995 Nov 10;270(45):27299–27304. doi: 10.1074/jbc.270.45.27299. [DOI] [PubMed] [Google Scholar]
  19. Kuipers O. P., Bierbaum G., Ottenwälder B., Dodd H. M., Horn N., Metzger J., Kupke T., Gnau V., Bongers R., van den Bogaard P. Protein engineering of lantibiotics. Antonie Van Leeuwenhoek. 1996 Feb;69(2):161–169. doi: 10.1007/BF00399421. [DOI] [PubMed] [Google Scholar]
  20. Kuipers O. P., Rollema H. S., Yap W. M., Boot H. J., Siezen R. J., de Vos W. M. Engineering dehydrated amino acid residues in the antimicrobial peptide nisin. J Biol Chem. 1992 Dec 5;267(34):24340–24346. [PubMed] [Google Scholar]
  21. Lian L. Y., Chan W. C., Morley S. D., Roberts G. C., Bycroft B. W., Jackson D. Solution structures of nisin A and its two major degradation products determined by n.m.r. Biochem J. 1992 Apr 15;283(Pt 2):413–420. doi: 10.1042/bj2830413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Liu W., Hansen J. N. The antimicrobial effect of a structural variant of subtilin against outgrowing Bacillus cereus T spores and vegetative cells occurs by different mechanisms. Appl Environ Microbiol. 1993 Feb;59(2):648–651. doi: 10.1128/aem.59.2.648-651.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Molenaar D., Abee T., Konings W. N. Continuous measurement of the cytoplasmic pH in Lactococcus lactis with a fluorescent pH indicator. Biochim Biophys Acta. 1991 Nov 14;1115(1):75–83. doi: 10.1016/0304-4165(91)90014-8. [DOI] [PubMed] [Google Scholar]
  24. Moll G. N., Roberts G. C., Konings W. N., Driessen A. J. Mechanism of lantibiotic-induced pore-formation. Antonie Van Leeuwenhoek. 1996 Feb;69(2):185–191. doi: 10.1007/BF00399423. [DOI] [PubMed] [Google Scholar]
  25. Moll G., Ubbink-Kok T., Hildeng-Hauge H., Nissen-Meyer J., Nes I. F., Konings W. N., Driessen A. J. Lactococcin G is a potassium ion-conducting, two-component bacteriocin. J Bacteriol. 1996 Feb;178(3):600–605. doi: 10.1128/jb.178.3.600-605.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morris S. L., Walsh R. C., Hansen J. N. Identification and characterization of some bacterial membrane sulfhydryl groups which are targets of bacteriostatic and antibiotic action. J Biol Chem. 1984 Nov 10;259(21):13590–13594. [PubMed] [Google Scholar]
  27. Nissen-Meyer J., Holo H., Håvarstein L. S., Sletten K., Nes I. F. A novel lactococcal bacteriocin whose activity depends on the complementary action of two peptides. J Bacteriol. 1992 Sep;174(17):5686–5692. doi: 10.1128/jb.174.17.5686-5692.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Okereke A., Montville T. J. Nisin dissipates the proton motive force of the obligate anaerobe Clostridium sporogenes PA 3679. Appl Environ Microbiol. 1992 Aug;58(8):2463–2467. doi: 10.1128/aem.58.8.2463-2467.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Reisinger P., Seidel H., Tschesche H., Hammes W. P. The effect of nisin on murein synthesis. Arch Microbiol. 1980 Oct;127(3):187–193. doi: 10.1007/BF00427192. [DOI] [PubMed] [Google Scholar]
  30. Ruhr E., Sahl H. G. Mode of action of the peptide antibiotic nisin and influence on the membrane potential of whole cells and on cytoplasmic and artificial membrane vesicles. Antimicrob Agents Chemother. 1985 May;27(5):841–845. doi: 10.1128/aac.27.5.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sahl H. G., Jack R. W., Bierbaum G. Biosynthesis and biological activities of lantibiotics with unique post-translational modifications. Eur J Biochem. 1995 Jun 15;230(3):827–853. doi: 10.1111/j.1432-1033.1995.tb20627.x. [DOI] [PubMed] [Google Scholar]
  32. Sahl H. G., Kordel M., Benz R. Voltage-dependent depolarization of bacterial membranes and artificial lipid bilayers by the peptide antibiotic nisin. Arch Microbiol. 1987;149(2):120–124. doi: 10.1007/BF00425076. [DOI] [PubMed] [Google Scholar]
  33. Van Den Hooven H. W., Doeland C. C., Van De Kamp M., Konings R. N., Hilbers C. W., Van De Ven F. J. Three-dimensional structure of the lantibiotic nisin in the presence of membrane-mimetic micelles of dodecylphosphocholine and of sodium dodecylsulphate. Eur J Biochem. 1996 Jan 15;235(1-2):382–393. doi: 10.1111/j.1432-1033.1996.00382.x. [DOI] [PubMed] [Google Scholar]
  34. Van Den Hooven H. W., Spronk C. A., Van De Kamp M., Konings R. N., Hilbers C. W., Van De Van F. J. Surface location and orientation of the lantibiotic nisin bound to membrane-mimicking micelles of dodecylphosphocholine and of sodium dodecylsulphate. Eur J Biochem. 1996 Jan 15;235(1-2):394–403. doi: 10.1111/j.1432-1033.1996.00394.x. [DOI] [PubMed] [Google Scholar]
  35. Van de Ven F. J., Van den Hooven H. W., Konings R. N., Hilbers C. W. NMR studies of lantibiotics. The structure of nisin in aqueous solution. Eur J Biochem. 1991 Dec 18;202(3):1181–1188. doi: 10.1111/j.1432-1033.1991.tb16488.x. [DOI] [PubMed] [Google Scholar]
  36. van den Hooven H. W., Fogolari F., Rollema H. S., Konings R. N., Hilbers C. W., van de Ven F. J. NMR and circular dichroism studies of the lantibiotic nisin in non-aqueous environments. FEBS Lett. 1993 Mar 15;319(1-2):189–194. doi: 10.1016/0014-5793(93)80065-3. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES