Application of a chemical probe to detect neutrophil elastase activation during inflammatory bowel disease

Beth Anderson, Daniel P. Poole, Luigi Aurelio, Garrett Zhen Ng, Markus Fleischmann, Paulina Kasperkiewicz, Celine Morissette, Marcin Drag, Ian R. van Driel, Brian L. Schmidt, Stephen J. Vanner, Nigel Bunnett, Laura Edgington-Mitchell

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Neutrophil elastase is a serine protease that has been implicated in the pathogenesis of inflammatory bowel disease. Due to post-translational control of its activation and high expression of its inhibitors in the gut, measurements of total expression poorly reflect the pool of active, functional neutrophil elastase. Fluorogenic substrate probes have been used to measure neutrophil elastase activity, though these tools lack specificity and traceability. PK105 is a recently described fluorescent activity-based probe, which binds to neutrophil elastase in an activity-dependent manner. The irreversible nature of this probe allows for accurate identification of its targets in complex protein mixtures. We describe the reactivity profile of PK105b, a new analogue of PK105, against recombinant serine proteases and in tissue extracts from healthy mice and from models of inflammation induced by oral cancer and Legionella pneumophila infection. We apply PK105b to measure neutrophil elastase activation in an acute model of experimental colitis. Neutrophil elastase activity is detected in inflamed, but not healthy, colons. We corroborate this finding in mucosal biopsies from patients with ulcerative colitis. Thus, PK105b facilitates detection of neutrophil elastase activity in tissue lysates, and we have applied it to demonstrate that this protease is unequivocally activated during colitis.

Original languageEnglish
Article number13295
Number of pages12
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 1 Dec 2019

Cite this

Anderson, B., Poole, D. P., Aurelio, L., Ng, G. Z., Fleischmann, M., Kasperkiewicz, P., ... Edgington-Mitchell, L. (2019). Application of a chemical probe to detect neutrophil elastase activation during inflammatory bowel disease. Scientific Reports, 9(1), [13295]. https://doi.org/10.1038/s41598-019-49840-4
Anderson, Beth ; Poole, Daniel P. ; Aurelio, Luigi ; Ng, Garrett Zhen ; Fleischmann, Markus ; Kasperkiewicz, Paulina ; Morissette, Celine ; Drag, Marcin ; van Driel, Ian R. ; Schmidt, Brian L. ; Vanner, Stephen J. ; Bunnett, Nigel ; Edgington-Mitchell, Laura. / Application of a chemical probe to detect neutrophil elastase activation during inflammatory bowel disease. In: Scientific Reports. 2019 ; Vol. 9, No. 1.
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abstract = "Neutrophil elastase is a serine protease that has been implicated in the pathogenesis of inflammatory bowel disease. Due to post-translational control of its activation and high expression of its inhibitors in the gut, measurements of total expression poorly reflect the pool of active, functional neutrophil elastase. Fluorogenic substrate probes have been used to measure neutrophil elastase activity, though these tools lack specificity and traceability. PK105 is a recently described fluorescent activity-based probe, which binds to neutrophil elastase in an activity-dependent manner. The irreversible nature of this probe allows for accurate identification of its targets in complex protein mixtures. We describe the reactivity profile of PK105b, a new analogue of PK105, against recombinant serine proteases and in tissue extracts from healthy mice and from models of inflammation induced by oral cancer and Legionella pneumophila infection. We apply PK105b to measure neutrophil elastase activation in an acute model of experimental colitis. Neutrophil elastase activity is detected in inflamed, but not healthy, colons. We corroborate this finding in mucosal biopsies from patients with ulcerative colitis. Thus, PK105b facilitates detection of neutrophil elastase activity in tissue lysates, and we have applied it to demonstrate that this protease is unequivocally activated during colitis.",
author = "Beth Anderson and Poole, {Daniel P.} and Luigi Aurelio and Ng, {Garrett Zhen} and Markus Fleischmann and Paulina Kasperkiewicz and Celine Morissette and Marcin Drag and {van Driel}, {Ian R.} and Schmidt, {Brian L.} and Vanner, {Stephen J.} and Nigel Bunnett and Laura Edgington-Mitchell",
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Anderson, B, Poole, DP, Aurelio, L, Ng, GZ, Fleischmann, M, Kasperkiewicz, P, Morissette, C, Drag, M, van Driel, IR, Schmidt, BL, Vanner, SJ, Bunnett, N & Edgington-Mitchell, L 2019, 'Application of a chemical probe to detect neutrophil elastase activation during inflammatory bowel disease', Scientific Reports, vol. 9, no. 1, 13295. https://doi.org/10.1038/s41598-019-49840-4

Application of a chemical probe to detect neutrophil elastase activation during inflammatory bowel disease. / Anderson, Beth; Poole, Daniel P.; Aurelio, Luigi; Ng, Garrett Zhen; Fleischmann, Markus; Kasperkiewicz, Paulina; Morissette, Celine; Drag, Marcin; van Driel, Ian R.; Schmidt, Brian L.; Vanner, Stephen J.; Bunnett, Nigel; Edgington-Mitchell, Laura.

In: Scientific Reports, Vol. 9, No. 1, 13295, 01.12.2019.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Anderson, Beth

AU - Poole, Daniel P.

AU - Aurelio, Luigi

AU - Ng, Garrett Zhen

AU - Fleischmann, Markus

AU - Kasperkiewicz, Paulina

AU - Morissette, Celine

AU - Drag, Marcin

AU - van Driel, Ian R.

AU - Schmidt, Brian L.

AU - Vanner, Stephen J.

AU - Bunnett, Nigel

AU - Edgington-Mitchell, Laura

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N2 - Neutrophil elastase is a serine protease that has been implicated in the pathogenesis of inflammatory bowel disease. Due to post-translational control of its activation and high expression of its inhibitors in the gut, measurements of total expression poorly reflect the pool of active, functional neutrophil elastase. Fluorogenic substrate probes have been used to measure neutrophil elastase activity, though these tools lack specificity and traceability. PK105 is a recently described fluorescent activity-based probe, which binds to neutrophil elastase in an activity-dependent manner. The irreversible nature of this probe allows for accurate identification of its targets in complex protein mixtures. We describe the reactivity profile of PK105b, a new analogue of PK105, against recombinant serine proteases and in tissue extracts from healthy mice and from models of inflammation induced by oral cancer and Legionella pneumophila infection. We apply PK105b to measure neutrophil elastase activation in an acute model of experimental colitis. Neutrophil elastase activity is detected in inflamed, but not healthy, colons. We corroborate this finding in mucosal biopsies from patients with ulcerative colitis. Thus, PK105b facilitates detection of neutrophil elastase activity in tissue lysates, and we have applied it to demonstrate that this protease is unequivocally activated during colitis.

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