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 language | English |
|---|---|
| Article number | 13295 |
| Number of pages | 12 |
| Journal | Scientific Reports |
| Volume | 9 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 1 Dec 2019 |
Cite this
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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 journal › Article › Research › peer-review
TY - JOUR
T1 - Application of a chemical probe to detect neutrophil elastase activation during inflammatory bowel disease
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
PY - 2019/12/1
Y1 - 2019/12/1
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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85072299634&partnerID=8YFLogxK
U2 - 10.1038/s41598-019-49840-4
DO - 10.1038/s41598-019-49840-4
M3 - Article
VL - 9
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 13295
ER -