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1.
Phys Ther Sport ; 65: 113-121, 2024 Jan.
Article in English | MEDLINE | ID: mdl-38128288

ABSTRACT

OBJECTIVES: To investigate the immediate effects of local muscle vibration (LMV) on static and dynamic balance control in individuals with and without chronic ankle instability (CAI). DESIGN: Quasi-experimental study. SETTING: Research laboratory. PARTICIPANTS: Twenty-six individuals with CAI and 26 healthy controls. MAIN OUTCOME MEASURES: Center of pressure variables (mean total velocity and displacement in anteroposterior (AP) and mediolateral (ML) directions) during single-leg standing with eyes open and eyes closed and also reach distances in anterior (ANT), posteromedial (PM), and posterolateral (PL) directions of the modified star excursion balance test (MSEBT) were assessed before and after LMV. RESULTS: Statistical analyses showed a significant decrease in mean total velocity and displacement in AP direction from before to after LMV in eyes open condition for both individuals with CAI (p = 0.025, p = 0.041, respectively) and healthy controls (p = 0.001, p = 0.003, respectively). Similar results were observed in eyes closed condition for both individuals with CAI (p < 0.001, p < 0.001, respectively) and healthy controls (p = 0.040, p = 0.014, respectively). The results also showed increased reach distances in ANT (p < 0.001), PM (p < 0.001), and PL directions (p < 0.001) in all participants after LMV. CONCLUSION: Our results suggest that LMV may be a useful tool in rehabilitation of static and dynamic balance deficits in individuals with CAI.


Subject(s)
Ankle , Joint Instability , Humans , Ankle Joint , Vibration/therapeutic use , Postural Balance/physiology , Muscles , Joint Instability/rehabilitation , Chronic Disease
2.
Am J Otolaryngol ; 44(4): 103911, 2023.
Article in English | MEDLINE | ID: mdl-37209538

ABSTRACT

OBJECTIVES: Muscle Tension Dysphonia is a voice disorder, which results in stiffness in the laryngeal extrinsic muscles, intense collision, painful contractions, and vibrations of the vocal cords. Due to the multifactorial identity of Muscle Tension Dysphonia, its therapeutic approach must be multidisciplinary. METHODS: The participants were divided into two groups: a control group (5participants) that received Circumlaryngeal Manual Therapy (CMT) + Placebo Transcutaneous Electrical Nerve Stimulation and an experimental group (5participants) that received Transcutaneous Electrical Nerve Stimulation + CMT. Both groups received 10 sessions of treatment, twice a week, for 40 min each. Before and after treatment, participants were assessed using the Dysphonia Severity Index (DSI) and surface electromyography for their ability to sustain the vowels /e& u/and count from 20 to30. RESULT: After therapy, there were substantial improvements in the DSI (2.72 ± 0.55, P < 0.05) and muscle electrical activity in the control group. The DSI (3.66 ± 0.63, P < 0.05) and muscle electrical activity were also significantly improved in the experimental group after treatment. The findings of the between-group comparison after treatment revealed a significantly greater increase in the Dysphonia Severity Index in the experimental group compared with the control group (P = 0.037). Although there was no significant difference between the two groups in terms of muscle electrical activity, clinically significant changes were more noticeable in the experimental group when compared with the control group. CONCLUSIONS: Positive results were seen in both groups. The results demonstrate that both approaches relax vocal tract muscles. As a result, Transcutaneous Electrical Nerve Stimulation was recommended as a complementary treatment for clients with Muscle Tension Dysphonia.


Subject(s)
Dysphonia , Transcutaneous Electric Nerve Stimulation , Humans , Dysphonia/therapy , Electromyography , Laryngeal Muscles , Muscle Tonus , Pilot Projects , Treatment Outcome , Voice Quality
3.
Physiother Theory Pract ; 39(2): 300-309, 2023 Feb.
Article in English | MEDLINE | ID: mdl-34983281

ABSTRACT

BACKGROUND: Proprioception deficit has been suggested as a possible mechanism contributing for the impaired postural control in low back pain (LBP) patients. Whether proprioception deficit is a result of or a cause of LBP has not been investigated. OBJECTIVE: The purpose of this study was to compare proprioceptive postural control strategies between prolonged standing induced low back pain developers (PDs) and non-pain developers (NPDs). METHOD: Thirty-two healthy subjects performed 1-h prolonged standing and their ratings of perceived LBP have been recorded. Eight quiet standing trials for 60 s performed immediately before and after the prolonged standing. Postural control was challenged by muscle vibration and different postural conditions during quiet standing. Data were recorded using a force platform. RESULTS: Forty percentage of participants is classified as PD. Before the prolonged standing, relative proprioceptive weighting was greater in the PD compared to NPD group (P = .029). Main effect of postural condition (F1,24 = 5.21, P = .032) and interaction of time by group (F1,24 = 8.08, P = .009) were significant for COP displacement in anteroposterior direction. Interaction of postural condition by group (F1,26 = 7.82, P = .010) and time by group (F1,26 = 9.71, P = .004) were significant for COP displacement in mediolateral direction. Main effect of postural condition (F1,26 = 6.31, P = .018) and interaction of postural condition by group (F1,26 = 7.07, P = .013) were significant for mean velocity in mediolateral direction. CONCLUSION: The PD group has altered proprioceptive postural control strategies before and after prolonged standing. Proprioception deficit should not be considered to be solely an adaptive response and may be causal for LBP development.


Subject(s)
Low Back Pain , Posture , Humans , Posture/physiology , Postural Balance/physiology , Proprioception/physiology , Standing Position
4.
Biochim Biophys Acta Proteins Proteom ; 1871(2): 140883, 2023 02 01.
Article in English | MEDLINE | ID: mdl-36455808

ABSTRACT

Multidrug resistance of bacteria and persistent infections related to biofilms, as well as the low availability of new antibacterial drugs, make it urgent to develop new antibiotics. Here, we evaluate the antibacterial and anti-biofilm properties of ticlopidine (TP), an anti-platelet aggregation drug, TP showed antibacterial activity against both gram-positive (MRSA) and gram-negative (E. coli, and P. aeruginosa) bacteria over a long treatment period. TP significantly reduced the survival of gram-negative bacteria in human blood though impact on gram-positives was more limited. TP may cause death in MRSA by inhibiting staphyloxanthin pigment synthesis, leading to oxidative stress, while scanning electron microscopy imaging indicate a loss of membrane integrity, damage, and consequent death due to lysis in gram-negative bacteria. TP showed good anti-biofilm activity against P. aeruginosa and MRSA, and a stronger biofilm degradation activity on P. aeruginosa compared to MRSA. Measuring fluorescence of the amyloid-reporter Thioflavin T (ThT) in biofilm implicated inhibition of amyloid formation as part of TP activity. This was confirmed by assays on the purified protein in P. aeruginosa, FapC, whose fibrillation kinetics was inhibited by TP. TP prolonged the lag phase of aggregation and reduced the subsequent growth rate and prolonging the lag phase to very long times provides ample opportunity to exert TP's antibacterial effect. We conclude that TP shows activity as an antibiotic against both gram-positive and gram-negative bacteria thanks to a broad range of activities, targeting bacterial metabolic processes, cellular structures and the biofilm matrix.


Subject(s)
Anti-Bacterial Agents , Escherichia coli , Humans , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Gram-Negative Bacteria/physiology , Gram-Positive Bacteria , Biofilms
5.
Molecules ; 27(13)2022 Jun 24.
Article in English | MEDLINE | ID: mdl-35807329

ABSTRACT

Functional amyloid is produced by many organisms but is particularly well understood in bacteria, where proteins such as CsgA (E. coli) and FapC (Pseudomonas) are assembled as functional bacterial amyloid (FuBA) on the cell surface in a carefully optimized process. Besides a host of helper proteins, FuBA formation is aided by multiple imperfect repeats which stabilize amyloid and streamline the aggregation mechanism to a fast-track assembly dominated by primary nucleation. These repeats, which are found in variable numbers in Pseudomonas, are most likely the structural core of the fibrils, though we still lack experimental data to determine whether the repeats give rise to ß-helix structures via stacked ß-hairpins (highly likely for CsgA) or more complicated arrangements (possibly the case for FapC). The response of FuBA fibrillation to denaturants suggests that nucleation and elongation involve equal amounts of folding, but protein chaperones preferentially target nucleation for effective inhibition. Smart peptides can be designed based on these imperfect repeats and modified with various flanking sequences to divert aggregation to less stable structures, leading to a reduction in biofilm formation. Small molecules such as EGCG can also divert FuBA to less organized structures, such as partially-folded oligomeric species, with the same detrimental effect on biofilm. Finally, the strong tendency of FuBA to self-assemble can lead to the formation of very regular two-dimensional amyloid films on structured surfaces such as graphite, which strongly implies future use in biosensors or other nanobiomaterials. In summary, the properties of functional amyloid are a much-needed corrective to the unfortunate association of amyloid with neurodegenerative disease and a testimony to nature's ability to get the best out of a protein fold.


Subject(s)
Escherichia coli , Neurodegenerative Diseases , Amyloid/chemistry , Amyloidogenic Proteins/chemistry , Bacterial Proteins/metabolism , Biofilms , Escherichia coli/metabolism , Humans , Pseudomonas/metabolism
6.
Int J Biol Macromol ; 209(Pt A): 426-440, 2022 Jun 01.
Article in English | MEDLINE | ID: mdl-35398391

ABSTRACT

The dangerous self-assembled and infectious seeds of α-synuclein (αSN) play primary roles in Parkinson's disease. Accordingly, the inhibition of αSN fibrillation and elimination of toxic aggregates are the main therapeutic strategies. Skullcapflavone II (S.FII), a compound isolated from S. pinnatifida, has shown multiple neuroprotective features. Herein, we demonstrated that S.FII inhibited αSN aggregation with IC50 of 7.2 µM. It increased nucleation time and decreased fibril elongation rate and the species formed in the presence of S.FII were unable to act as seeds. Additionally, S.FII inhibited both secondary nucleation and seeding of αSN and disaggregated the mature preformed fibrils as well. The species formed in the presence of S.FII showed less toxicity. It also preserved neurite length and dopamine content of SH-SY5Y cells and attenuated the inflammatory responses in mixed glial cells. The Localized Surface Plasmon Resonance (LSPR) analysis indicated that S.FII interacts with αSN. Docking simulation studies on αSN fibrils revealed that S.FII could interact with the key residues of the salt bridges and glutamine ladder, which might lead to the destruction of fibril's structures. We also showed that S.FII passes through the blood-brain barrier in vitro and in vivo. Overall, these findings elucidate the neuroprotective roles of S.FII in reducing αSN pathogenicity.


Subject(s)
Parkinson Disease , alpha-Synuclein , Flavonoids/pharmacology , Humans , alpha-Synuclein/chemistry
7.
Chem Sci ; 13(2): 536-553, 2022 Jan 05.
Article in English | MEDLINE | ID: mdl-35126986

ABSTRACT

Unlike misfolding in neurodegenerative diseases, aggregation of functional amyloids involved in bacterial biofilm, e.g. CsgA (E. coli) and FapC (Pseudomonas), is carefully regulated. However, it is unclear whether functional aggregation is inhibited by chaperones targeting pathological misfolding and if so by what mechanism. Here we analyze how four entirely different human chaperones or protein modulators (transthyretin, S100A9, Bri2 BRICHOS and DNAJB6) and bacterial CsgC affect CsgA and FapC fibrillation. CsgA is more susceptible to inhibition than FapC and the chaperones vary considerably in the efficiency of their inhibition. However, mechanistic analysis reveals that all predominantly target primary nucleation rather than elongation or secondary nucleation, while stoichiometric considerations suggest that DNAJB6 and CsgC target nuclei rather than monomers. Inhibition efficiency broadly scales with the chaperones' affinity for monomeric CsgA and FapC. The chaperones tend to target the most aggregation-prone regions of CsgA, but do not display such tendencies towards the more complex FapC sequence. Importantly, the most efficient inhibitors (Bri2 BRICHOS and DNAJB6) significantly reduce bacterial biofilm formation. This commonality of chaperone action may reflect the simplicity of functional amyloid formation, driven largely by primary nucleation, as well as the ability of non-bacterial chaperones to deploy their proteostatic capacities across biological kingdoms.

8.
BBA Adv ; 2: 100055, 2022.
Article in English | MEDLINE | ID: mdl-37082589

ABSTRACT

Functional amyloids (FA) are proteins which are evolutionarily optimized to form highly stable fibrillar structures that strengthen the bacterial biofilm matrix. FA such as CsgA (E. coli) and FapC (Pseudomonas) are secreted to the bacterial surface where they integrate into growing fibril structures projecting from the outer membrane. FA are exposed to membrane surfaces in this process, but it remains unclear how membranes can interact with FA and potentially affect the self-assembly. Here we report the effect of different vesicles (DOPG, DMPG, DOPS, DOPC and DMPC) on the kinetics and structural endpoints of FapC fibrillation using various biophysical techniques. Particularly anionic lipids such as DMPG trigger FapC fibrillation, and the protein's second repeat sequence (R2) appears to be important for this interaction. Vesicles formed from phospholipids extracted from three different Pseudomonas strains (Δfap, ΔFapC and pfap) induce FapC fibrillation by accelerating nucleation. The general aggregation inhibitor epigallocatechin gallate (EGCG) inhibits FapC fibrillation by blocking interactions between FapC and vesicles and redirecting FapC monomers to oligomer structures. Our work indicates that biological membranes can contribute significantly to the fibrillation of functional amyloid.

9.
Small Methods ; 5(6): e2001002, 2021 06.
Article in English | MEDLINE | ID: mdl-34927901

ABSTRACT

Formation of amyloid structures is originally linked to human disease. However, amyloid materials are found extensively in the animal and bacterial world where they stabilize intra- and extra-cellular environments like biofilms or cell envelopes. To date, functional amyloids have largely been studied using optical microscopy techniques in vivo, or after removal from their biological context for higher-resolution studies in vitro. Furthermore, conventional microscopies only indirectly identify amyloids based on morphology or unspecific amyloid dyes. Here, the high chemical and spatial (≈20 nm) resolution of Infrared Nanospectroscopy (AFM-IR) to investigate functional amyloid from Escherichia coli (curli), Pseudomonas (Fap), and the Archaea Methanosaeta (MspA) in situ is exploited. It is demonstrated that AFM-IR identifies amyloid protein within single intact cells through their cross ß-sheet secondary structure, which has a unique spectroscopic signature in the amide I band of protein. Using this approach, nanoscale-resolved chemical images and spectra of purified curli and Methanosaeta cell wall sheaths are provided. The results highlight significant differences in secondary structure between E. coli cells with and without curli. Taken together, these results suggest that AFM-IR is a new and powerful label-free tool for in situ investigations of the biophysical state of functional amyloid and biomolecules in general.


Subject(s)
Amyloid/isolation & purification , Amyloid/metabolism , Amyloidogenic Proteins/metabolism , Archaea/metabolism , Bacteria/metabolism , Amyloid/chemistry , Amyloidogenic Proteins/chemistry , Amyloidogenic Proteins/isolation & purification , Bacterial Outer Membrane , Biofilms , Escherichia coli/metabolism , Escherichia coli Proteins , Humans , Protein Structure, Secondary , Pseudomonas/metabolism
10.
Int J Mol Sci ; 22(21)2021 Oct 27.
Article in English | MEDLINE | ID: mdl-34769023

ABSTRACT

Aggregated α-synuclein (α-syn) is the main constituent of Lewy bodies, which are a pathological hallmark of Parkinson's disease (PD). Environmental factors are thought to be potential triggers capable of initiating the aggregation of the otherwise monomeric α-syn. Braak's seminal work redirected attention to the intestine and recent reports of dysbiosis have highlighted the potential causative role of the microbiome in the initiation of pathology of PD. Staphylococcus aureus is a bacterium carried by 30-70% of the general population. It has been shown to produce functional amyloids, called phenol soluble modulins (PSMαs). Here, we studied the kinetics of α-syn aggregation under quiescent conditions in the presence or absence of four different PSMα peptides and observed a remarkable shortening of the lag phase in their presence. Whereas pure α-syn monomer did not aggregate up to 450 h after initiation of the experiment in neither neutral nor mildly acidic buffer, the addition of different PSMα peptides resulted in an almost immediate increase in the Thioflavin T (ThT) fluorescence. Despite similar peptide sequences, the different PSMα peptides displayed distinct effects on the kinetics of α-syn aggregation. Kinetic analyses of the data suggest that all four peptides catalyze α-syn aggregation through heterogeneous primary nucleation. The immunogold electron microscopic analyses showed that the aggregates were fibrillar and composed of α-syn. In addition of the co-aggregated materials to a cell model expressing the A53T α-syn variant fused to GFP was found to catalyze α-syn aggregation and phosphorylation in the cells. Our results provide evidence of a potential trigger of synucleinopathies and could have implications for the prevention of the diseases.


Subject(s)
Phenols/metabolism , Protein Aggregation, Pathological/metabolism , Staphylococcus aureus/metabolism , alpha-Synuclein/metabolism , Amyloid , Cell Line , HEK293 Cells , Humans , Parkinson Disease/metabolism , Phosphorylation/physiology
11.
ACS Omega ; 6(34): 21960-21970, 2021 Aug 31.
Article in English | MEDLINE | ID: mdl-34497891

ABSTRACT

Functional amyloids are highly organized protein/peptide structures that inter alia promote biofilm formation in different bacteria. One such example is provided by a family of 20-45 residue-long peptides called phenol-soluble modulins (PSMs) from Staphylococcus aureus. External components such as eukaryotic host proteins, which alter self-assembly of bacterial amyloids, can affect the biofilm matrix. Here, we studied the effect of the highly prevalent human plasma protein fibrinogen (Fg) on fibrillation of PSMs. Fg inhibits or suppresses fibrillation of most PSMs tested (PSMα1, PSMß1, and PSMß2) except for PSMα3, whose already rapid aggregation is accelerated even further by Fg but leads to amorphous ß-rich aggregates rather than fibrils. Fg also induces PSMß2 to form amorphous aggregates and diverts PSMα1 into off-pathway oligomers which consist of both Fg and PSMα1 and cannot seed fibrillation. Peptide arrays showed that Fg bound to the N-terminus of PSMα1, while it bound to the entire length of PSMα3 (except the C terminus) and to the C-termini of PSMß1 and PSMß2. The latter peptides are all positively charged, while Fg is negatively charged at physiological pH. The positive charges complement Fg's net negative charge of -7.6 at pH 7.4. Fg's ability to inhibit PSM fibrillation reveals a potential host-defense mechanism to prevent bacterial biofilm growth and infections in the human body.

12.
J Biol Chem ; 297(2): 100953, 2021 08.
Article in English | MEDLINE | ID: mdl-34270957

ABSTRACT

Phenol-soluble modulins (PSMs), such as α-PSMs, ß-PSMs, and δ-toxin, are virulence peptides secreted by different Staphylococcus aureus strains. PSMs are able to form amyloid fibrils, which may strengthen the biofilm matrix that promotes bacterial colonization of and extended growth on surfaces (e.g., cell tissue) and increases antibiotic resistance. Many components contribute to biofilm formation, including the human-produced highly sulfated glycosaminoglycan heparin. Although heparin promotes S. aureus infection, the molecular basis for this is unclear. Given that heparin is known to induce fibrillation of a wide range of proteins, we hypothesized that heparin aids bacterial colonization by promoting PSM fibrillation. Here, we address this hypothesis using a combination of thioflavin T-fluorescence kinetic studies, CD, FTIR, electron microscopy, and peptide microarrays to investigate the mechanism of aggregation, the structure of the fibrils, and identify possible binding regions. We found that heparin accelerates fibrillation of all α-PSMs (except PSMα2) and δ-toxin but inhibits ß-PSM fibrillation by blocking nucleation or reducing fibrillation levels. Given that S. aureus secretes higher levels of α-PSM than ß-PSM peptides, heparin is therefore likely to promote fibrillation overall. Heparin binding is driven by multiple positively charged lysine residues in α-PSMs and δ-toxins, the removal of which strongly reduced binding affinity. Binding of heparin did not affect the structure of the resulting fibrils, that is, the outcome of the aggregation process. Rather, heparin provided a scaffold to catalyze or inhibit fibrillation. Based on our findings, we speculate that heparin may strengthen the bacterial biofilm and therefore enhance colonization via increased PSM fibrillation.


Subject(s)
Peptides , Staphylococcus aureus , Bacterial Toxins , Biofilms/growth & development , Kinetics , Peptides/metabolism , Virulence
13.
Front Microbiol ; 11: 573253, 2020.
Article in English | MEDLINE | ID: mdl-33240231

ABSTRACT

Staphylococcus aureus is a human pathogen that can cause chronic and recurrent infections and is recalcitrant to antibiotic chemotherapy. This trait is partly attributed to its ability to form persister cells, which are subpopulations of cells that are tolerant to lethal concentrations of antibiotics. Recently, we showed that the phenol-soluble modulins (PSMs) expressed by S. aureus reduce persister cell formation. PSMs are a versatile group of toxins that, in addition to toxicity, form amyloid-like fibrils thought to support biofilm structures. Here, we examined individual or combined synthetic PSMα peptides and their equivalent amyloid-like fibrils on ciprofloxacin-selected S. aureus persister cells. We found that PSMα2 and the mixture of all four alpha peptides consistently were able to reduce persister frequency in all growth phases, and this activity was specifically linked to the presence of the soluble peptide as no effect was seen with fibrillated peptides. Persister reduction was particularly striking in a mutant that, due to mutations in the Krebs cycle, has enhanced ability to form persisters with PSMα4 and the combination of peptides being most effective. In biofilms, only the combination of peptides displayed persister reducing activity. Collectively, we report the individual contributions of PSMα peptides to persister cell reduction and that the combination of peptides generally was most effective. Strikingly, the fibrillated peptides lost activity and thus, if formed in bacterial cultures, they will be inactive against persister cells. Further studies will be needed to address the biological role of phenol-soluble modulins in reducing persister cells.

14.
Biomolecules ; 9(11)2019 10 26.
Article in English | MEDLINE | ID: mdl-31717821

ABSTRACT

Self-assembly of proteins to ß-sheet rich amyloid fibrils is commonly observed in various neurodegenerative diseases. However, amyloid also occurs in the extracellular matrix of bacterial biofilm, which protects bacteria from environmental stress and antibiotics. Many Pseudomonas strains produce functional amyloid where the main component is the highly fibrillation-prone protein FapC. FapC fibrillation may be inhibited by small molecules such as plant polyphenols, which are already known to inhibit formation of pathogenic amyloid, but the mechanism and biological impact of inhibition is unclear. Here, we elucidate how polyphenols modify the self-assembly of functional amyloid, with particular focus on epigallocatechin gallate (EGCG), penta-O-galloyl-ß-d-glucose (PGG), baicalein, oleuropein, and procyanidin B2. We find EGCG and PGG to be the best inhibitors. These compounds inhibit amyloid formation by redirecting the aggregation of FapC monomers into oligomeric species, which according to small-angle X-ray scattering (SAXS) measurements organize into core-shell complexes of short axis diameters 25-26 nm consisting of ~7 monomers. Using peptide arrays, we identify EGCG-binding sites in FapC's linker regions, C and N-terminal parts, and high amyloidogenic sequences located in the R2 and R3 repeats. We correlate our biophysical observations to biological impact by demonstrating that the extent of amyloid inhibition by the different inhibitors correlated with their ability to reduce biofilm, highlighting the potential of anti-amyloid polyphenols as therapeutic agents against biofilm infections.


Subject(s)
Amyloid/metabolism , Catechin/analogs & derivatives , Fungal Proteins/metabolism , Hydrolyzable Tannins/pharmacology , Polyphenols/pharmacology , Pseudomonas/drug effects , Amyloid/genetics , Biofilms/drug effects , Catechin/pharmacology , Fungal Proteins/genetics , Protein Aggregates/drug effects , Pseudomonas/physiology
15.
Biochim Biophys Acta Proteins Proteom ; 1867(11): 140263, 2019 11.
Article in English | MEDLINE | ID: mdl-31421227

ABSTRACT

Pseudomonas species export the amyloid-forming protein FapC to strengthen bacterial biofilm. P. species also produce the biosurfactant rhamnolipid (Rhl) and its outer membrane contains lipopolysaccharide (LPS). Given the possible contacts between FapC, Rhl and LPS, we here investigate how Rhl and LPS affect FapC fibrillation compared with SDS, known to promote fibrillation of proteins at sub-micellar concentrations. Micelles of all three surfactants help FapC bypass the nucleation lag phase, leading to rapid fibrillation, which persists even at high concentrations of micelles and incorporates almost all available FapC monomers. Fibrils formed at high micellar concentrations of Rhl and SDS seed fibrillation at low surfactant concentrations while retaining the original fibril structure. FapC interacts strongly with SDS to form a dense network of narrow fibrils. Small angle X-ray scattering (SAXS) analyses reveal that surfactants reduce the population of intermediates in the fibrillation process and detect a fast aggregation step over the first 2-4 h which precedes the main fibrillation monitored by Thioflavin T. An additional SAXS-detected rearrangement of early aggregates occurs after 4-10 h. At high Rhl concentrations, the micelles are decorated with protein fibrils. SDS induces FapC fibrillation so efficiently that epigallocatechin-3-gallate (EGCG) is unable to inhibit this process. However, EGCG stimulates FapC oligomer formation and inhibits fibrillation both on its own and in the presence of Rhl and LPS. This oligomer could be modelled as a compact core with a flexible shell. This suggests that EGCG can override the natural amyloid-stimulatory properties of these biosurfactants and thus target biofilm.


Subject(s)
Amyloidogenic Proteins/chemistry , Bacterial Proteins/chemistry , Glycolipids/chemistry , Lipopolysaccharides/chemistry , Protein Aggregates , Pseudomonas aeruginosa/chemistry
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