Abstract
Lactococcin G is a novel lactococcal bacteriocin whose activity depends on the complementary action of two peptides, termed alpha and beta. Peptide synthesis of the alpha and beta peptides yielded biologically active lactococcin G, which was used in mode-of-action studies on sensitive cells of Lactococcus lactis. Approximately equivalent amounts of both peptides were required for optimal bactericidal effect. No effect was observed with either the alpha or beta peptide in the absence of the complementary peptide. The combination of alpha and beta peptides (lactococcin G) dissipates the membrane potential (delta omega), and as a consequence cells release alpha-aminoisobutyrate, a non-metabolizable alanine analog that is accumulated through a proton motive-force dependent mechanism. In addition, the cellular ATP level is dramatically reduced, which results in a drastic decrease of the ATP-driven glutamate uptake. Lactococcin G does not form a proton-conducting pore, as it has no effect on the transmembrane pH gradient. Dissipation of the membrane potential by uncouplers causes a slow release of potassium (rubidium) ions. However, rapid release of potassium was observed in the presence of lactococcin G. These data suggest that the bactericidal effect of lactococcin G is due to the formation of potassium-selective channels by the alpha and beta peptides in the target bacterial membrane.
Full Text
The Full Text of this article is available as a PDF (238.7 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abee T., Klaenhammer T. R., Letellier L. Kinetic studies of the action of lactacin F, a bacteriocin produced by Lactobacillus johnsonii that forms poration complexes in the cytoplasmic membrane. Appl Environ Microbiol. 1994 Mar;60(3):1006–1013. doi: 10.1128/aem.60.3.1006-1013.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bakker E. P., Harold F. M. Energy coupling to potassium transport in Streptococcus faecalis. Interplay of ATP and the protonmotive force. J Biol Chem. 1980 Jan 25;255(2):433–440. [PubMed] [Google Scholar]
- 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]
- Chikindas M. L., García-Garcerá M. J., Driessen A. J., Ledeboer A. M., Nissen-Meyer J., Nes I. F., Abee T., Konings W. N., Venema G. Pediocin PA-1, a bacteriocin from Pediococcus acidilactici PAC1.0, forms hydrophilic pores in the cytoplasmic membrane of target cells. Appl Environ Microbiol. 1993 Nov;59(11):3577–3584. doi: 10.1128/aem.59.11.3577-3584.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chopin A., Chopin M. C., Moillo-Batt A., Langella P. Two plasmid-determined restriction and modification systems in Streptococcus lactis. Plasmid. 1984 May;11(3):260–263. doi: 10.1016/0147-619x(84)90033-7. [DOI] [PubMed] [Google Scholar]
- Driessen A. J., Kodde J., de Jong S., Konings W. N. Neutral amino acid transport by membrane vesicles of Streptococcus cremoris is subject to regulation by internal pH. J Bacteriol. 1987 Jun;169(6):2748–2754. doi: 10.1128/jb.169.6.2748-2754.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Driessen A. J., de Vrij W., Konings W. N. Functional incorporation of beef-heart cytochrome c oxidase into membranes of Streptococcus cremoris. Eur J Biochem. 1986 Feb 3;154(3):617–624. doi: 10.1111/j.1432-1033.1986.tb09443.x. [DOI] [PubMed] [Google Scholar]
- Driessen A. J., de Vrij W., Konings W. N. Incorporation of beef heart cytochrome c oxidase as a proton-motive force-generating mechanism in bacterial membrane vesicles. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7555–7559. doi: 10.1073/pnas.82.22.7555. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Jiménez-Díaz R., Rios-Sánchez R. M., Desmazeaud M., Ruiz-Barba J. L., Piard J. C. Plantaricins S and T, Two New Bacteriocins Produced by Lactobacillus plantarum LPCO10 Isolated from a Green Olive Fermentation. Appl Environ Microbiol. 1993 May;59(5):1416–1424. doi: 10.1128/aem.59.5.1416-1424.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- 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]
- Nes I. F., Havarstein L. S., Holo H. Genetics of non-lantibiotic bacteriocins. Dev Biol Stand. 1995;85:645–651. [PubMed] [Google Scholar]
- 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]
- Nissen-Meyer J., Larsen A. G., Sletten K., Daeschel M., Nes I. F. Purification and characterization of plantaricin A, a Lactobacillus plantarum bacteriocin whose activity depends on the action of two peptides. J Gen Microbiol. 1993 Sep;139(9):1973–1978. doi: 10.1099/00221287-139-9-1973. [DOI] [PubMed] [Google Scholar]
- Ojcius D. M., Young J. D. Cytolytic pore-forming proteins and peptides: is there a common structural motif? Trends Biochem Sci. 1991 Jun;16(6):225–229. doi: 10.1016/0968-0004(91)90090-i. [DOI] [PubMed] [Google Scholar]
- Poolman B., Hellingwerf K. J., Konings W. N. Regulation of the glutamate-glutamine transport system by intracellular pH in Streptococcus lactis. J Bacteriol. 1987 May;169(5):2272–2276. doi: 10.1128/jb.169.5.2272-2276.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poolman B., Molenaar D., Smid E. J., Ubbink T., Abee T., Renault P. P., Konings W. N. Malolactic fermentation: electrogenic malate uptake and malate/lactate antiport generate metabolic energy. J Bacteriol. 1991 Oct;173(19):6030–6037. doi: 10.1128/jb.173.19.6030-6037.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poolman B., Smid E. J., Konings W. N. Kinetic properties of a phosphate-bond-driven glutamate-glutamine transport system in Streptococcus lactis and Streptococcus cremoris. J Bacteriol. 1987 Jun;169(6):2755–2761. doi: 10.1128/jb.169.6.2755-2761.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Skaugen M., Nissen-Meyer J., Jung G., Stevanovic S., Sletten K., Inger C., Abildgaard M., Nes I. F. In vivo conversion of L-serine to D-alanine in a ribosomally synthesized polypeptide. J Biol Chem. 1994 Nov 4;269(44):27183–27185. [PubMed] [Google Scholar]
- Stoffels G., Guthmundsdóttir A., Abee T. Membrane-associated proteins encoded by the nisin gene cluster may function as a receptor for the lantibiotic carnocin UI49. Microbiology. 1994 Jun;140(Pt 6):1443–1450. doi: 10.1099/00221287-140-6-1443. [DOI] [PubMed] [Google Scholar]
- Venema K., Abee T., Haandrikman A. J., Leenhouts K. J., Kok J., Konings W. N., Venema G. Mode of Action of Lactococcin B, a Thiol-Activated Bacteriocin from Lactococcus lactis. Appl Environ Microbiol. 1993 Apr;59(4):1041–1048. doi: 10.1128/aem.59.4.1041-1048.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zajdel J. K., Ceglowski P., Dobrazański W. T. Mechanism of action of lactostrepcin 5, a bacteriocin produced by Streptococcus cremoris 202. Appl Environ Microbiol. 1985 Apr;49(4):969–974. doi: 10.1128/aem.49.4.969-974.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Belkum M. J., Hayema B. J., Jeeninga R. E., Kok J., Venema G. Organization and nucleotide sequences of two lactococcal bacteriocin operons. Appl Environ Microbiol. 1991 Feb;57(2):492–498. doi: 10.1128/aem.57.2.492-498.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Belkum M. J., Kok J., Venema G., Holo H., Nes I. F., Konings W. N., Abee T. The bacteriocin lactococcin A specifically increases permeability of lactococcal cytoplasmic membranes in a voltage-independent, protein-mediated manner. J Bacteriol. 1991 Dec;173(24):7934–7941. doi: 10.1128/jb.173.24.7934-7941.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]