1887

Abstract

The development of antibacterial resistance is inevitable and is a major concern in hospitals and communities. Moreover, biofilm-grown bacteria are less sensitive to antimicrobial treatment. In this respect, the Gram-positive is an important source of nosocomial biofilm-associated infections. In the search for new antibacterial therapies, the type I signal peptidase (SPase I) serves as a potential target for development of antibacterials with a novel mode of action. This enzyme cleaves off the signal peptide from secreted proteins, making it essential for protein secretion, and hence for bacterial cell viability. encodes three putative SPases I (denoted Sip1, Sip2 and Sip3), of which Sip1 lacks the catalytic lysine. In this report, we investigated the active SPases I in more detail. Sip2 and Sip3 were found to complement a temperature-sensitive mutant, demonstrating their functional activity. functional activity of purified Sip2 and Sip3 proteins and inhibition of their activity by the SPase I inhibitor arylomycin A were further illustrated using a fluorescence resonance energy transfer (FRET)-based assay. Furthermore, we demonstrated that SPase I not only is an attractive target for development of novel antibacterials against free-living bacteria, but also is a feasible target for biofilm-associated infections.

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2009-11-01
2020-01-21
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References

  1. Allsop, A., Brooks, G., Edwards, P. D., Kaura, A. C. & Southgate, R. ( 1996; ). Inhibitors of bacterial signal peptidase: a series of 6-(substituted oxyethyl)penems. J Antibiot (Tokyo) 49, 921–928.[CrossRef]
    [Google Scholar]
  2. Black, M. T. ( 1993; ). Evidence that the catalytic activity of prokaryote leader peptidase depends upon the operation of a serine–lysine catalytic dyad. J Bacteriol 175, 4957–4961.
    [Google Scholar]
  3. Black, M. T. & Bruton, G. ( 1998; ). Inhibitors of bacterial signal peptidases. Curr Pharm Des 4, 133–154.
    [Google Scholar]
  4. Bockstael, K., Geukens, N., Rao, C. V., Herdewijn, P., Anne, J. & Van Aerschot, A. ( 2009; ). An easy and fast method for the evaluation of Staphylococcus epidermidis type I signal peptidase inhibitors. J Microbiol Methods 78, 231–237.[CrossRef]
    [Google Scholar]
  5. Bruton, G., Huxley, A., O'Hanlon, P., Orlek, B., Eggleston, D., Humphries, J., Readshaw, S., West, A., Ashman, S. & other authors ( 2003; ). Lipopeptide substrates for SpsB, the Staphylococcus aureus type I signal peptidase: design, conformation and conversion to α-ketoamide inhibitors. Eur J Med Chem 38, 351–356.[CrossRef]
    [Google Scholar]
  6. Christensen, G. D., Simpson, W. A., Bisno, A. L. & Beachey, E. H. ( 1982; ). Adherence of slime-producing strains of Staphylococcus epidermidis to smooth surfaces. Infect Immun 37, 318–326.
    [Google Scholar]
  7. Christensen, G. D., Simpson, W. A., Younger, J. J., Baddour, L. M., Barrett, F. F., Melton, D. M. & Beachey, E. H. ( 1985; ). Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol 22, 996–1006.
    [Google Scholar]
  8. Cohen, M. L. ( 2000; ). Changing patterns of infectious disease. Nature 406, 762–767.[CrossRef]
    [Google Scholar]
  9. Cregg, K. M., Wilding, I. & Black, M. T. ( 1996; ). Molecular cloning and expression of the spsB gene encoding an essential type I signal peptidase from Staphylococcus aureus. J Bacteriol 178, 5712–5718.
    [Google Scholar]
  10. Dalbey, R. E., Lively, M. O., Bron, S. & van Dijl, J. M. ( 1997; ). The chemistry and enzymology of the type I signal peptidases. Protein Sci 6, 1129–1138.[CrossRef]
    [Google Scholar]
  11. Donlan, R. M. & Costerton, J. W. ( 2002; ). Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15, 167–193.[CrossRef]
    [Google Scholar]
  12. Geukens, N., Lammertyn, E., Van Mellaert, L., Schacht, S., Schaerlaekens, K., Parro, V., Bron, S., Engelborghs, Y., Mellado, R. P. & Anne, J. ( 2001; ). Membrane topology of the Streptomyces lividans type I signal peptidases. J Bacteriol 183, 4752–4760.[CrossRef]
    [Google Scholar]
  13. Geukens, N., Lammertyn, E., Van Mellaert, L., Engelborghs, Y., Mellado, R. P. & Anne, J. ( 2002; ). Physical requirements for in vitro processing of the Streptomyces lividans signal peptidases. J Biotechnol 96, 79–91.[CrossRef]
    [Google Scholar]
  14. Hall-Stoodley, L., Costerton, J. W. & Stoodley, P. ( 2004; ). Bacterial biofilms: from the natural environment to infectious diseases. Nat Rev Microbiol 2, 95–108.[CrossRef]
    [Google Scholar]
  15. Inada, T., Court, D. L., Ito, K. & Nakamura, Y. ( 1989; ). Conditionally lethal amber mutations in the leader peptidase gene of Escherichia coli. J Bacteriol 171, 585–587.
    [Google Scholar]
  16. Kavanaugh, J. S., Thoendel, M. & Horswill, A. R. ( 2007; ). A role for type I signal peptidase in Staphylococcus aureus quorum sensing. Mol Microbiol 65, 780–798.[CrossRef]
    [Google Scholar]
  17. Kulanthaivel, P., Kreuzman, A. J., Strege, M. A., Belvo, M. D., Smitka, T. A., Clemens, M., Swartling, J. R., Minton, K. L., Zheng, F. & other authors ( 2004; ). Novel lipoglycopeptides as inhibitors of bacterial signal peptidase I. J Biol Chem 279, 36250–36258.[CrossRef]
    [Google Scholar]
  18. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.[CrossRef]
    [Google Scholar]
  19. Lammertyn, E., Van Mellaert, L., Meyen, E., Lebeau, I., De Buck, E., Anne, J. & Geukens, N. ( 2004; ). Molecular and functional characterization of type I signal peptidase from Legionella pneumophila. Microbiology 150, 1475–1483.[CrossRef]
    [Google Scholar]
  20. Li, X., Yan, Z. & Xu, J. ( 2003; ). Quantitative variation of biofilms among strains in natural populations of Candida albicans. Microbiology 149, 353–362.[CrossRef]
    [Google Scholar]
  21. Mack, D., Siemssen, N. & Laufs, R. ( 1992; ). Parallel induction by glucose of adherence and a polysaccharide antigen specific for plastic-adherent Staphylococcus epidermidis: evidence for functional relation to intercellular adhesion. Infect Immun 60, 2048–2057.
    [Google Scholar]
  22. Miller, J. H. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  23. Musial-Siwek, M., Kendall, D. A. & Yeagle, P. L. ( 2008; ). Solution NMR of signal peptidase, a membrane protein. Biochim Biophys Acta 1778, 937–944.[CrossRef]
    [Google Scholar]
  24. NCCLS ( 2003; ). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically: approved standard M7–A6, 6th edn. Wayne, PA: National Committee for Clinical Laboratory Standards.
  25. Novick, R. P. ( 1991; ). Genetic systems in staphylococci. Methods Enzymol 204, 587–636.
    [Google Scholar]
  26. O'Brien, J., Wilson, I., Orton, T. & Pognan, F. ( 2000; ). Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem 267, 5421–5426.[CrossRef]
    [Google Scholar]
  27. O'Gara, J. P. & Humphreys, H. ( 2001; ). Staphylococcus epidermidis biofilms: importance and implications. J Med Microbiol 50, 582–587.
    [Google Scholar]
  28. Paetzel, M., Dalbey, R. E. & Strynadka, N. C. ( 1998; ). Crystal structure of a bacterial signal peptidase in complex with a β-lactam inhibitor. Nature 396, 186–190.[CrossRef]
    [Google Scholar]
  29. Paetzel, M., Dalbey, R. E. & Strynadka, N. C. ( 2002; ). Crystal structure of a bacterial signal peptidase apoenzyme: implications for signal peptide binding and the Ser–Lys dyad mechanism. J Biol Chem 277, 9512–9519.[CrossRef]
    [Google Scholar]
  30. Paetzel, M., Goodall, J. J., Kania, M., Dalbey, R. E. & Page, M. G. ( 2004; ). Crystallographic and biophysical analysis of a bacterial signal peptidase in complex with a lipopeptide-based inhibitor. J Biol Chem 279, 30781–30790.[CrossRef]
    [Google Scholar]
  31. Pettit, R. K., Weber, C. A., Kean, M. J., Hoffmann, H., Pettit, G. R., Tan, R., Franks, K. S. & Horton, M. L. ( 2005; ). Microplate Alamar Blue assay for Staphylococcus epidermidis biofilm susceptibility testing. Antimicrob Agents Chemother 49, 2612–2617.[CrossRef]
    [Google Scholar]
  32. Pitts, B., Hamilton, M. A., Zelver, N. & Stewart, P. S. ( 2003; ). A microtiter-plate screening method for biofilm disinfection and removal. J Microbiol Methods 54, 269–276.[CrossRef]
    [Google Scholar]
  33. Roberts, T. C., Smith, P. A., Cirz, R. T. & Romesberg, F. E. ( 2007; ). Structural and initial biological analysis of synthetic arylomycin A2. J Am Chem Soc 129, 15830–15838.[CrossRef]
    [Google Scholar]
  34. Romesberg, F. E. ( 2009; ). Re-evaluating Discarded Antibiotic Candidates. Abstracts of the Society for General Microbiology Spring Meeting, Harrogate, UK, 30 March–2 April 2009. http://www.sgm.ac.uk/meetings/pdfabstracts/harrogate2009abs.pdf
  35. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  36. Schulin, T. & Voss, A. ( 2001; ). Coagulase-negative staphylococci as a cause of infections related to intravascular prosthetic devices: limitations of present therapy. Clin Microbiol Infect 7 (Suppl. 4), 1–7.
    [Google Scholar]
  37. Strathmann, M., Wingender, J. & Flemming, H. C. ( 2002; ). Application of fluorescently labelled lectins for the visualization and biochemical characterization of polysaccharides in biofilms of Pseudomonas aeruginosa. J Microbiol Methods 50, 237–248.[CrossRef]
    [Google Scholar]
  38. Studier, F. W. & Moffatt, B. A. ( 1986; ). Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol 189, 113–130.[CrossRef]
    [Google Scholar]
  39. Studier, F. W., Rosenberg, A. H., Dunn, J. J. & Dubendorff, J. W. ( 1990; ). Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol 185, 60–89.
    [Google Scholar]
  40. Sutherland, I. ( 2001; ). Biofilm exopolysaccharides: a strong and sticky framework. Microbiology 147, 3–9.
    [Google Scholar]
  41. Tschantz, W. R., Sung, M., Delgado-Partin, V. M. & Dalbey, R. E. ( 1993; ). A serine and a lysine residue implicated in the catalytic mechanism of the Escherichia coli leader peptidase. J Biol Chem 268, 27349–27354.
    [Google Scholar]
  42. Vandecasteele, S. J., Peetermans, W. E., Merckx, R., Van Ranst, M. & Van Eldere, J. ( 2002; ). Use of gDNA as internal standard for gene expression in staphylococci in vitro and in vivo. Biochem Biophys Res Commun 291, 528–534.[CrossRef]
    [Google Scholar]
  43. van Roosmalen, M. L., Geukens, N., Jongbloed, J. D., Tjalsma, H., Dubois, J. Y., Bron, S., van Dijl, J. M. & Anne, J. ( 2004; ). Type I signal peptidases of Gram-positive bacteria. Biochim Biophys Acta 1694, 279–297.[CrossRef]
    [Google Scholar]
  44. Villain-Guillot, P., Gualtieri, M., Bastide, L. & Leonetti, J. P. ( 2007; ). In vitro activities of different inhibitors of bacterial transcription against Staphylococcus epidermidis biofilm. Antimicrob Agents Chemother 51, 3117–3121.[CrossRef]
    [Google Scholar]
  45. Yoneyama, H. & Katsumata, R. ( 2006; ). Antibiotic resistance in bacteria and its future for novel antibiotic development. Biosci Biotechnol Biochem 70, 1060–1075.[CrossRef]
    [Google Scholar]
  46. Zhang, Y. Q., Ren, S. X., Li, H. L., Wang, Y. X., Fu, G., Yang, J., Qin, Z. Q., Miao, Y. G., Wang, W. Y. & other authors ( 2003; ). Genome-based analysis of virulence genes in a non-biofilm-forming Staphylococcus epidermidis strain (ATCC 12228). Mol Microbiol 49, 1577–1593.[CrossRef]
    [Google Scholar]
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