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Thrombin-derived C-terminal fragments aggregate and scavenge bacteria and their proinflammatory products

Petrlova, Jitka LU ; Petruk, Ganna LU ; Huber, Roland G. ; McBurnie, Eilish W. ; van der Plas, Mariena J.A. LU ; Bond, Peter J. ; Puthia, Manoj LU and Schmidtchen, Artur LU (2020) In Journal of Biological Chemistry 295(11). p.3417-3430
Abstract

Thrombin-derived C-terminal peptides (TCPs), including a major 11-kDa fragment (TCP96), are produced through cleavage by human neutrophil elastase and aggregate lipopolysaccharide (LPS) and the Gram-negative bacterium Escherichia coli. However, the physiological roles of TCP96 in controlling bacterial infections and reducing LPS-induced inflammation are unclear. Here, using various biophysical methods, in silico molecular modeling, microbiological and cellular assays, and animal models, we examined the structural features and functional roles of recombinant TCP96 (rTCP96) in the aggregation of multiple bacteria and the Toll-like receptor (TLR) agonists they produce. We found that rTCP96 aggregates both Gram-negative and Gram-positive... (More)

Thrombin-derived C-terminal peptides (TCPs), including a major 11-kDa fragment (TCP96), are produced through cleavage by human neutrophil elastase and aggregate lipopolysaccharide (LPS) and the Gram-negative bacterium Escherichia coli. However, the physiological roles of TCP96 in controlling bacterial infections and reducing LPS-induced inflammation are unclear. Here, using various biophysical methods, in silico molecular modeling, microbiological and cellular assays, and animal models, we examined the structural features and functional roles of recombinant TCP96 (rTCP96) in the aggregation of multiple bacteria and the Toll-like receptor (TLR) agonists they produce. We found that rTCP96 aggregates both Gram-negative and Gram-positive bacteria, including Staphylococcus aureus and Pseudomonas aeruginosa, and their cell-wall components LPS, lipid A, and lipoteichoic acid (LTA). The Gram-negative bacteria E. coli and P. aeruginosa were particularly sensitive to aggregation-induced bacterial permeabilization and killing. As a proof of concept, we show that rTCP96 reduces LPS-induced NF-κB activation in human monocytes, as well as in mouse models of LPS-induced subcutaneous inflammation. Moreover, in a mouse model of subcutaneous inoculation with P. aeruginosa, rTCP96 reduced bacterial levels. Together, these results link TCP-mediated aggregation of endotoxins and bacteria in vitro to attenuation of inflammation and bacterial levels in vivo.

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Contribution to journal
publication status
published
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in
Journal of Biological Chemistry
volume
295
issue
11
pages
14 pages
publisher
ASBMB
external identifiers
  • pmid:32034093
  • scopus:85081971719
ISSN
0021-9258
DOI
10.1074/jbc.RA120.012741
language
English
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yes
id
e3a1c99a-0662-4f3b-adc1-57c663d861a9
date added to LUP
2020-04-06 13:19:57
date last changed
2020-04-07 05:45:17
@article{e3a1c99a-0662-4f3b-adc1-57c663d861a9,
  abstract     = {<p>Thrombin-derived C-terminal peptides (TCPs), including a major 11-kDa fragment (TCP96), are produced through cleavage by human neutrophil elastase and aggregate lipopolysaccharide (LPS) and the Gram-negative bacterium Escherichia coli. However, the physiological roles of TCP96 in controlling bacterial infections and reducing LPS-induced inflammation are unclear. Here, using various biophysical methods, in silico molecular modeling, microbiological and cellular assays, and animal models, we examined the structural features and functional roles of recombinant TCP96 (rTCP96) in the aggregation of multiple bacteria and the Toll-like receptor (TLR) agonists they produce. We found that rTCP96 aggregates both Gram-negative and Gram-positive bacteria, including Staphylococcus aureus and Pseudomonas aeruginosa, and their cell-wall components LPS, lipid A, and lipoteichoic acid (LTA). The Gram-negative bacteria E. coli and P. aeruginosa were particularly sensitive to aggregation-induced bacterial permeabilization and killing. As a proof of concept, we show that rTCP96 reduces LPS-induced NF-κB activation in human monocytes, as well as in mouse models of LPS-induced subcutaneous inflammation. Moreover, in a mouse model of subcutaneous inoculation with P. aeruginosa, rTCP96 reduced bacterial levels. Together, these results link TCP-mediated aggregation of endotoxins and bacteria in vitro to attenuation of inflammation and bacterial levels in vivo.</p>},
  author       = {Petrlova, Jitka and Petruk, Ganna and Huber, Roland G. and McBurnie, Eilish W. and van der Plas, Mariena J.A. and Bond, Peter J. and Puthia, Manoj and Schmidtchen, Artur},
  issn         = {0021-9258},
  language     = {eng},
  number       = {11},
  pages        = {3417--3430},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Thrombin-derived C-terminal fragments aggregate and scavenge bacteria and their proinflammatory products},
  url          = {http://dx.doi.org/10.1074/jbc.RA120.012741},
  doi          = {10.1074/jbc.RA120.012741},
  volume       = {295},
  year         = {2020},
}