Daniel Otzen

TY - JOUR

T1 - Epigallocatechin Gallate Remodels overexpressed Functional Amyloids in Pseudomonas aeruginosa and Increases Biofilm Susceptibility to Antibiotic Treatment

AU - Stenvang, Marcel

AU - Dueholm, Morten S

AU - Vad, Brian S

AU - Seviour, Thomas W

AU - Zeng, Guanghong

AU - Geifman-Shochat, Susana

AU - Søndergaard, Mads T

AU - Christiansen, Gunna

AU - Meyer, Rikke Louise

AU - Kjelleberg, Staffan

AU - Otzen, Daniel E

AU - Nielsen, Per Halkjær

N1 - Copyright © 2016, The American Society for Biochemistry and Molecular Biology.

PY - 2016/12/16

Y1 - 2016/12/16

N2 - Epigallocatechin-3-gallate (EGCG) is the major polyphenol in green tea. It has anti-microbial properties and disrupts the ordered structure of amyloid fibrils involved in human disease. The anti-microbial effect of EGCG against the opportunistic pathogen Pseudomonas aeruginosa has been shown to involve disruption of quorum sensing (QS). Functional amyloid fibrils in P. aeruginosa (Fap) are able to bind and retain quorum sensing molecules, suggesting that EGCG interferes with QS through structural remodeling of amyloid fibrils. Here we show that EGCG inhibits the ability of Fap to form fibrils; instead, EGCG stabilizes protein oligomers. Existing fibrils are remodeled by EGCG into non-amyloid aggregates. This fibril remodeling increases the binding of pyocyanin, demonstrating a mechanism by which EGCG can affect the QS function of functional amyloid. EGCG reduced the amyloid specific fluorescent Thioflavin T signal in P. aeruginosa biofilms at concentrations known to have exert an antimicrobial effect. Nanoindentation studies showed that EGCG reduced the stiffness of biofilm containing Fap fibrils but not in biofilm with little Fap. In a combination treatment with EGCG and tobramycin, EGCG had a moderate effect on the minimum bactericidal eradication concentration against wildtype P. aeruginosa biofilms while EGCG had a more pronounced effect when Fap were overexpressed. Our results provide a direct molecular explanation for EGCG's ability to disrupt P. aeruginosa's QS and modify its biofilm, and strengthens the case for EGCG as a candidate in multidrug treatment of persistent biofilm infections.

AB - Epigallocatechin-3-gallate (EGCG) is the major polyphenol in green tea. It has anti-microbial properties and disrupts the ordered structure of amyloid fibrils involved in human disease. The anti-microbial effect of EGCG against the opportunistic pathogen Pseudomonas aeruginosa has been shown to involve disruption of quorum sensing (QS). Functional amyloid fibrils in P. aeruginosa (Fap) are able to bind and retain quorum sensing molecules, suggesting that EGCG interferes with QS through structural remodeling of amyloid fibrils. Here we show that EGCG inhibits the ability of Fap to form fibrils; instead, EGCG stabilizes protein oligomers. Existing fibrils are remodeled by EGCG into non-amyloid aggregates. This fibril remodeling increases the binding of pyocyanin, demonstrating a mechanism by which EGCG can affect the QS function of functional amyloid. EGCG reduced the amyloid specific fluorescent Thioflavin T signal in P. aeruginosa biofilms at concentrations known to have exert an antimicrobial effect. Nanoindentation studies showed that EGCG reduced the stiffness of biofilm containing Fap fibrils but not in biofilm with little Fap. In a combination treatment with EGCG and tobramycin, EGCG had a moderate effect on the minimum bactericidal eradication concentration against wildtype P. aeruginosa biofilms while EGCG had a more pronounced effect when Fap were overexpressed. Our results provide a direct molecular explanation for EGCG's ability to disrupt P. aeruginosa's QS and modify its biofilm, and strengthens the case for EGCG as a candidate in multidrug treatment of persistent biofilm infections.

U2 - 10.1074/jbc.M116.739953

DO - 10.1074/jbc.M116.739953

M3 - Journal article

C2 - 27784787

VL - 291

SP - 26540

EP - 26553

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 51

ER -