scholarly journals Strain-Dependent Adsorption of Pseudomonas aeruginosa-Derived Adhesin-like Peptides at Abiotic Surfaces

Micro ◽  
2021 ◽  
Vol 1 (1) ◽  
pp. 129-139
Author(s):  
Yu Yang ◽  
Sabrina Schwiderek ◽  
Guido Grundmeier ◽  
Adrian Keller
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Molecular Level ◽  
Microbial Colonization ◽  
Peptide Sequence ◽  
Type Iv ◽  
Research Activities ◽  
Abiotic Surfaces ◽  
Peptide Adsorption ◽  
Pilin Protein ◽  
Strain Dependent

Implant-associated infections are an increasingly severe burden on healthcare systems worldwide and many research activities currently focus on inhibiting microbial colonization of biomedically relevant surfaces. To obtain molecular-level understanding of the involved processes and interactions, we investigate the adsorption of synthetic adhesin-like peptide sequences derived from the type IV pili of the Pseudomonas aeruginosa strains PAK and PAO at abiotic model surfaces, i.e., Au, SiO2, and oxidized Ti. These peptides correspond to the sequences of the receptor-binding domain 128–144 of the major pilin protein, which is known to facilitate P. aeruginosa adhesion at biotic and abiotic surfaces. Using quartz crystal microbalance with dissipation monitoring (QCM-D), we find that peptide adsorption is material- as well as strain-dependent. At the Au surface, PAO(128–144) shows drastically stronger adsorption than PAK(128–144), whereas adsorption of both peptides is markedly reduced at the oxide surfaces with less drastic differences between the two sequences. These observations suggest that peptide adsorption is influenced by not only the peptide sequence, but also peptide conformation. Our results furthermore highlight the importance of molecular-level investigations to understand and ultimately control microbial colonization of surfaces.

The Pseudomonas aeruginosa type IV pilin receptor binding domain functions as an adhesin for both biotic and abiotic surfaces

2006 ◽  
Vol 60 (3) ◽  
pp. 813-813 ◽  
Author(s):  
Carmen L. Giltner ◽  
Erin J. van Schaik ◽  
Gerald F. Audette ◽  
Dan Kao ◽  
Robert S. Hodges ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Receptor Binding ◽  
Binding Domain ◽  
Receptor Binding Domain ◽  
Type Iv ◽  
Abiotic Surfaces ◽  
Type Iv Pilin

scholarly journals A Geobacter sulfurreducens Strain Expressing Pseudomonas aeruginosa Type IV Pili Localizes OmcS on Pili but Is Deficient in Fe(III) Oxide Reduction and Current Production

2013 ◽  
Vol 80 (3) ◽  
pp. 1219-1224 ◽  
Author(s):  
Xing Liu ◽  
Pier-Luc Tremblay ◽  
Nikhil S. Malvankar ◽  
Kelly P. Nevin ◽  
Derek R. Lovley ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Electron Transfer ◽  
Structural Features ◽  
Geobacter Sulfurreducens ◽  
Control Strain ◽  
Oxide Reduction ◽  
Content Type ◽  
Type Iv ◽  
Current Production ◽  
Pilin Protein

ABSTRACTThe conductive pili ofGeobacterspecies play an important role in electron transfer to Fe(III) oxides, in long-range electron transport through current-producing biofilms, and in direct interspecies electron transfer. Although multiple lines of evidence have indicated that the pili ofGeobacter sulfurreducenshave a metal-like conductivity, independent of the presence ofc-type cytochromes, this claim is still controversial. In order to further investigate this phenomenon, a strain ofG. sulfurreducens, designated strain PA, was constructed in which the gene for the native PilA, the structural pilin protein, was replaced with the PilA gene ofPseudomonas aeruginosaPAO1. Strain PA expressed and properly assembledP. aeruginosaPilA subunits into pili and exhibited a profile of outer surfacec-type cytochromes similar to that of a control strain expressing theG. sulfurreducensPilA. Surprisingly, the strain PA pili were decorated with thec-type cytochrome OmcS in a manner similar to the control strain. However, the strain PA pili were 14-fold less conductive than the pili of the control strain, and strain PA was severely impaired in Fe(III) oxide reduction and current production. These results demonstrate that the presence of OmcS on pili is not sufficient to confer conductivity to pili and suggest that there are unique structural features of theG. sulfurreducensPilA that are necessary for conductivity.

scholarly journals Pseudomonas aeruginosa Type IV Pilus Expression in Neisseria gonorrhoeae: Effects of Pilin Subunit Composition on Function and Organelle Dynamics

2007 ◽  
Vol 189 (18) ◽  
pp. 6676-6685 ◽  
Author(s):  
Hanne C. Winther-Larsen ◽  
Matthew C. Wolfgang ◽  
Jos P. M. van Putten ◽  
Norbert Roos ◽  
Finn Erik Aas ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Neisseria Gonorrhoeae ◽  
Type Iv Pili ◽  
Type Iv ◽  
Pilin Subunit ◽  
Steady State Levels ◽  
Natural Genetic Transformation ◽  
Necessary And Sufficient ◽  
Pilin Protein ◽  
Behavior Sensitivity

ABSTRACT Type IV pili (TFP) play central roles in the expression of many phenotypes including motility, multicellular behavior, sensitivity to bacteriophages, natural genetic transformation, and adherence. In Neisseria gonorrhoeae, these properties require ancillary proteins that act in conjunction with TFP expression and influence organelle dynamics. Here, the intrinsic contributions of the pilin protein itself to TFP dynamics and associated phenotypes were examined by expressing the Pseudomonas aeruginosa PilAPAK pilin subunit in N. gonorrhoeae. We show here that, although PilAPAK pilin can be readily assembled into TFP in this background, steady-state levels of purifiable fibers are dramatically reduced relative those of endogenous pili. This defect is due to aberrant TFP dynamics as it is suppressed in the absence of the PilT pilus retraction ATPase. Functionally, PilAPAK pilin complements gonococcal adherence for human epithelial cells but only in a pilT background, and this property remains dependent on the coexpression of both the PilC adhesin and the PilV pilin-like protein. Since P. aeruginosa pilin only moderately supports neisserial sequence-specific transformation despite its assembly proficiency, these results together suggest that PilAPAK pilin functions suboptimally in this environment. This appears to be due to diminished compatibility with resident proteins essential for TFP function and dynamics. Despite this, PilAPAK pili support retractile force generation in this background equivalent to that reported for endogenous pili. Furthermore, PilAPAK pili are both necessary and sufficient for bacteriophage PO4 binding, although the strain remains phage resistant. Together, these findings have significant implications for TFP biology in both N. gonorrhoeae and P. aeruginosa.

scholarly journals The Pseudomonas aeruginosa type IV pilin receptor binding domain functions as an adhesin for both biotic and abiotic surfaces

2006 ◽  
Vol 59 (4) ◽  
pp. 1083-1096 ◽  
Author(s):  
Carmen L. Giltner ◽  
Erin J. van Schaik ◽  
Gerald F. Audette ◽  
Dan Kao ◽  
Robert S. Hodges ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Receptor Binding ◽  
Binding Domain ◽  
Receptor Binding Domain ◽  
Type Iv ◽  
Abiotic Surfaces ◽  
Type Iv Pilin

scholarly journals Dual-Function Vaccine for Pseudomonas aeruginosa: Characterization of Chimeric Exotoxin A-Pilin Protein

2001 ◽  
Vol 69 (11) ◽  
pp. 6962-6969 ◽  
Author(s):  
Ralf Hertle ◽  
Randall Mrsny ◽  
David J. Fitzgerald
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Chimeric Protein ◽  
Infectious Agent ◽  
Bacterial Adherence ◽  
Dual Function ◽  
Exotoxin A ◽  
Type Iv ◽  
Type Iv Pilin ◽  
Immunological Assays ◽  
Pilin Protein

ABSTRACT Pseudomonas aeruginosa is the major infectious agent of concern for cystic fibrosis patients. Strategies to prevent colonization by this bacterium and/or neutralize its virulence factors are clearly needed. Here we characterize a dual-function vaccine designed to generate antibodies to reduce bacterial adherence and to neutralize the cytotoxic activity of exotoxin A. To construct the vaccine, key sequences from type IV pilin were inserted into a vector encoding a nontoxic (active-site deletion) version of exotoxin A. The chimeric protein, termed PE64Δ553pil, was expressed inEscherichia coli, refolded to a near-native conformation, and then characterized by various biochemical and immunological assays. PE64Δ553pil bound specifically to asialo-GM1, and, when injected into rabbits, produced antibodies that reduced bacterial adherence and neutralized the cell-killing activity of exotoxin A. Results support further evaluation of this chimeric protein as a vaccine to prevent Pseudomonascolonization in susceptible individuals.

scholarly journals Transcriptional Activation of the tad Type IVb Pilus Operon by PypB in Yersinia enterocolitica

2010 ◽  
Vol 192 (14) ◽  
pp. 3809-3821 ◽  
Author(s):  
Jennifer Schilling ◽  
Karin Wagner ◽  
Stephanie Seekircher ◽  
Lilo Greune ◽  
Verena Humberg ◽  
...  
Keyword(s):  
Yersinia Enterocolitica ◽  
Transcriptional Activation ◽  
Genetic Linkage ◽  
Genomic Island ◽  
Transcriptional Regulator ◽  
Type Iv Pili ◽  
Type Iv ◽  
Abiotic Surfaces ◽  
Archaeal Species ◽  
Pilin Protein

ABSTRACT Type IV pili are virulence factors in various bacteria and mediate, among other functions, the colonization of diverse surfaces. Various subclasses of type IV pili have been identified, but information on pilus expression, biogenesis, and the associated phenotypes is sparse for the genus Yersinia. We recently described the identification of PypB as a transcriptional regulator in Yersinia enterocolitica. Here we show that the pypB gene is associated with the tad locus, a genomic island that is widespread among bacterial and archaeal species. The genetic linkage of pypB with the tad locus is conserved throughout the yersiniae but is not found among other bacteria carrying the tad locus. We show that the genes of the tad locus form an operon in Y. enterocolitica that is controlled by PypB and that pypB is part of this operon. The tad genes encode functions necessary for the biogenesis of the Flp subfamily of type IVb pili initially described for Aggregatibacter actinomycetemcomitans to mediate a tight-adherence phenotype. In Y. enterocolitica, the Flp pilin protein shows some peculiarities in its amino acid sequence that imply similarities as well as differences compared to typical motifs found in the Flp subtype of type IVb pili. Flp is expressed and processed after PypB overproduction, resulting in microcolony formation but not in increased adherence to biotic or abiotic surfaces. Our data describe the transcriptional regulation of the tad type IVb pilus operon by PypB in Y. enterocolitica but fail to show most previously described phenotypes associated with this type of pilus in other bacteria.

scholarly journals Sub-Inhibitory Concentrations of Ciprofloxacin Alone and Combinations with Plant-Derived Compounds against P. aeruginosa Biofilms and Their Effects on the Metabolomic Profile of P. aeruginosa Biofilms

Antibiotics ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 414
Author(s):  
Didem Kart ◽  
Tuba Reçber ◽  
Emirhan Nemutlu ◽  
Meral Sagiroglu
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Inhibitory Concentration ◽  
Molecular Level ◽  
Metabolomic Profile ◽  
Gene Expressions ◽  
New Agents ◽  
Qrt Pcr ◽  
Metabolomic Approach

Introduction: Alternative anti-biofilm agents are needed to combat Pseudomonas aeruginosa infections. The mechanisms behind these new agents also need to be revealed at a molecular level. Materials and methods: The anti-biofilm effects of 10 plant-derived compounds on P. aeruginosa biofilms were investigated using minimum biofilm eradication concentration (MBEC) and virulence assays. The effects of ciprofloxacin and compound combinations on P. aeruginosa in mono and triple biofilms were compared. A metabolomic approach and qRT-PCR were applied to the biofilms treated with ciprofloxacin in combination with baicalein, esculin hydrate, curcumin, and cinnamaldehyde at sub-minimal biofilm inhibitory concentration (MBIC) concentrations to highlight the specific metabolic shifts between the biofilms and to determine the quorum sensing gene expressions, respectively. Results: The combinations of ciprofloxacin with curcumin, baicalein, esculetin, and cinnamaldehyde showed more reduced MBICs than ciprofloxacin alone. The quorum sensing genes were downregulated in the presence of curcumin and cinnamaldehyde, while upregulated in the presence of baicalein and esculin hydrate rather than for ciprofloxacin alone. The combinations exhibited different killing effects on P. aeruginosa in mono and triple biofilms without affecting its virulence. The findings of the decreased metabolite levels related to pyrimidine and lipopolysaccharide synthesis and to down-regulated alginate and lasI expressions strongly indicate the role of multifactorial mechanisms for curcumin-mediated P. aeruginosa growth inhibition. Conclusions: The use of curcumin, baicalein, esculetin, and cinnamaldehyde with ciprofloxacin will help fight against P. aeruginosa biofilms. To the best of our knowledge, this is the first study of its kind to define the effect of plant-based compounds as possible anti-biofilm agents with low MBICs for the treatment of P. aeruginosa biofilms through metabolomic pathways.

scholarly journals Aztreonam Lysine Increases the Activity of Phages E79 and phiKZ against Pseudomonas aeruginosa PA01

2021 ◽  
Vol 9 (1) ◽  
pp. 152
Author(s):  
Carly M. Davis ◽  
Jaclyn G. McCutcheon ◽  
Jonathan J. Dennis
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Morphological Changes ◽  
Burst Size ◽  
Type Iv Pili ◽  
Biofilm Growth ◽  
Promising Alternative ◽  
Type Iv ◽  
Alternative Treatment Option ◽  
One Step ◽  
Step Growth

Pseudomonas aeruginosa is a pernicious bacterial pathogen that is difficult to treat because of high levels of antibiotic resistance. A promising alternative treatment option for such bacteria is the application of bacteriophages; the correct combination of phages plus antibiotics can produce synergistic inhibitory effects. In this study, we describe morphological changes induced by sub-MIC levels of the antibiotic aztreonam lysine (AzLys) on P. aeruginosa PA01, which may in part explain the observed phage–antibiotic synergy (PAS). One-step growth curves for phage E79 showed increased adsorption rates, decreased infection latency, accelerated time to lysis and a minor reduction in burst size. Phage E79 plus AzLys PAS was also able to significantly reduce P. aeruginosa biofilm growth over 3-fold as compared to phage treatment alone. Sub-inhibitory AzLys-induced filamentation of P. aeruginosa cells resulted in loss of twitching motility and a reduction in swimming motility, likely due to a reduction in the number of polar Type IV pili and flagella, respectively, on the filamented cell surfaces. Phage phiKZ, which uses Type IV pili as a receptor, did not exhibit increased activity with AzLys at lower sub-inhibitory levels, but still produced phage–antibiotic synergistic killing with sub-inhibitory AzLys. A one-step growth curve indicates that phiKZ in the presence of AzLys also exhibits a decreased infection latency and moderately undergoes accelerated time to lysis. In contrast to prior PAS studies demonstrating that phages undergo delayed time to lysis with cell filamentation, these PAS results show that phages undergo accelerated time to lysis, which therefore suggests that PAS is dependent upon multiple factors, including the type of phages and antibiotics used, and the bacterial host being tested.

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