Use of luminescent bacteria for rapid screening and characterization of short cationic antimicrobial peptides synthesized on cellulose using peptide array technology

Hilpert, Kai; Hancock, Robert E. W.

doi:10.1038/nprot.2007.203

Cited by 71 publications

(61 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Assays on the detrimental effect of high eDNA concentrations on strains of S. oneidensis MR-1 were performed as described previously (22,28,40). Overnight cultures of the appropriate strains carrying pBBR1-MCS5-TT-Pmot-lux were reinoculated into fresh medium and were grown to the mid-exponential-growth phase.…”

Section: Methodsmentioning

confidence: 99%

Functional Specificity of Extracellular Nucleases of Shewanella oneidensis MR-1

Heun

Binnenkade

Kreienbaum

et al. 2012

Appl Environ Microbiol

View full text Add to dashboard Cite

ABSTRACTBacterial species such asShewanella oneidensisMR-1 require extracellular nucleolytic activity for the utilization of extracellular DNA (eDNA) as a source of nutrients and for the turnover of eDNA as a structural matrix component during biofilm formation. We have previously characterized two extracellular nucleases ofS. oneidensisMR-1, ExeM and ExeS. Although both are involved in biofilm formation, they are not specifically required for the utilization of eDNA as a nutrient. Here we identified and characterized EndA, a third extracellular nuclease ofShewanella. The heterologously overproduced and purified protein was highly active and rapidly degraded linear and supercoiled DNAs of various origins. Divalent metal ions (Mg2+or Mn2+) were required for function.endAis cotranscribed withphoA, an extracellular phosphatase, and is not upregulated upon phosphostarvation. Deletion ofendAabolished both extracellular degradation of DNA byS. oneidensisMR-1 and the ability to use eDNA as a sole source of phosphorus. PhoA is not strictly required for the exploitation of eDNA as a nutrient. The activity of EndA prevents the formation of large cell aggregates during planktonic growth. However, in contrast to the findings for ExeM,endAdeletion had only minor effects on biofilm formation. The findings strongly suggest that the extracellular nucleases ofS. oneidensisexert specific functions required under different conditions.

show abstract

Section: Methodsmentioning

confidence: 99%

Functional Specificity of Extracellular Nucleases of Shewanella oneidensis MR-1

Heun

Binnenkade

Kreienbaum

et al. 2012

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…13). This strategy was applied to identify regulatory sequences or promoter regions in the genome (70,71) and in drug screening (72)(73)(74).…”

Section: Applicationsmentioning

confidence: 99%

Firefly bioluminescence: A mechanistic approach of luciferase catalyzed reactions

2008

View full text Add to dashboard Cite

SummaryLuciferase is a general term for enzymes catalyzing visible light emission by living organisms (bioluminescence). The studies carried out with Photinus pyralis (firefly) luciferase allowed the discovery of the reaction leading to light production. It can be regarded as a two-step process: the first corresponds to the reaction of luciferase's substrate, luciferin (LH 2 ), with ATPMg 21 generating inorganic pyrophosphate and an intermediate luciferyl-adenylate (LH 2 -AMP); the second is the oxidation and decarboxylation of LH 2 -AMP to oxyluciferin, the light emitter, producing CO 2 , AMP, and photons of yellow-green light (550-570 nm). In a dark reaction LH 2 -AMP is oxidized to dehydroluciferyl-adenylate (L-AMP). Luciferase also shows acyl-coenzyme A synthetase activity, which leads to the formation of dehydroluciferyl-coenzyme A (L-CoA), luciferyl-coenzyme A (LH 2 -CoA), and fatty acyl-CoAs. Moreover luciferase catalyzes the synthesis of dinucleoside polyphosphates from nucleosides with at least a 3 0 -phosphate chain plus an intact terminal pyrophosphate moiety. The LH 2 stereospecificity is a particular feature of the bioluminescent reaction where each isomer, D-LH 2 or L-LH 2 , has a specific function. Practical applications of the luciferase system, either in its native form or with engineered proteins, encloses the analytical assay of metabolites like ATP and molecular biology studies with luc as a reporter gene, including the most recent and increasing field of bioimaging.2008 IUBMB IUBMB Life, 61(1):

show abstract

“…Our understanding of the amino acid sequence requirements that underlie the antimicrobial activities of CAMPs has been accelerated by the development of high-throughput assays and methods to analyze their physiochemical characteristics (18)(19)(20). Physiochemical characteristics of amino acids have been defined by dozens of quantitative structure-activity relationships (QSARs), including various descriptors of hydrophobicity, hydrophilicity, isoelectric point, charge, mass, volume, and other physiochemical characteristics (21).…”

mentioning

confidence: 99%

Targeting Mycobacterium tuberculosis and Other Microbial Pathogens Using Improved Synthetic Antibacterial Peptides

Ramón-García

Mikut

et al. 2013

Antimicrob Agents Chemother

Self Cite

View full text Add to dashboard Cite

The lack of effective therapies for treating tuberculosis (TB) is a global health problem. While Mycobacterium tuberculosis is notoriously resistant to most available antibiotics, we identified synthetic short cationic antimicrobial peptides that were active at low micromolar concentrations (less than 10 M). These small peptides (averaging 10 amino acids) had remarkably broad spectra of antimicrobial activities against both bacterial and fungal pathogens and an indication of low cytotoxicity. In addition, their antimicrobial activities displayed various degrees of species specificity that were not related to taxonomy. For example, Candida albicans and Staphylococcus aureus were the best surrogates to predict peptide activity against M. tuberculosis, while Mycobacterium smegmatis was a poor surrogate. Principle component analysis of activity spectrum profiles identified unique features associated with activity against M. tuberculosis that reflect their distinctive amino acid composition; active peptides were more hydrophobic and cationic, reflecting increased tryptophan with compensating decreases in valine and other uncharged amino acids and increased lysine. These studies provide foundations for development of cationic antimicrobial peptides as potential new therapeutic agents for TB treatment.

show abstract

Use of luminescent bacteria for rapid screening and characterization of short cationic antimicrobial peptides synthesized on cellulose using peptide array technology

Cited by 71 publications

References 21 publications

Functional Specificity of Extracellular Nucleases of Shewanella oneidensis MR-1

Functional Specificity of Extracellular Nucleases of Shewanella oneidensis MR-1

Firefly bioluminescence: A mechanistic approach of luciferase catalyzed reactions

Targeting Mycobacterium tuberculosis and Other Microbial Pathogens Using Improved Synthetic Antibacterial Peptides

Contact Info

Product

Resources

About