Measuring A. alternata Protease Activity and Their Effects on Asthma

Persistent Link:
http://hdl.handle.net/10150/297595
Title:
Measuring A. alternata Protease Activity and Their Effects on Asthma
Author:
Graham, Lauren Victoria; Flynn, Andrea N.; Sherwood, Cara L.; Schultz, Stephanie M.; Hoffman, Justin; Daines, Michael O.; Boitano, Scott
Issue Date:
2013
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
Alternaria alternata, is a key factor in childhood allergic asthma in semi-arid climates. We have shown that filtrates from A. alternata can both cause asthma symptoms in a mouse model as well as directly activate human bronchial epithelial cells, which provide the first point of contact for microbes in the airway lumen. These interactions are dependent on A. alternata protease activity within the host acting on protease activated receptor-2 (PAR₂) (1). Proteolysis of PAR₂ results in an exposed “tethered ligand” that initiates host intracellular signaling pathways and subsequent physiological response (1). In this report, we measured the protease activity of various A. alternata filtrate samples using a standard, casein-based fluorescent protease detection assay and a novel fluorescent probe we designed to include the specific proteolytic sequence of PAR₂. Both assays showed protease activity of A. alternata filtrates that matched their respective signaling responses in host epithelial cells. Conditions that limited protease activity effectively blocked protease detections in the assays and cell signaling responses. We conclude that A. alternata filtrates induce airway epithelial cell signaling and asthmatic response partly via protease activity on PAR₂. By utilizing our novel proteolytic assay we may better predict potency of various PAR₂-dependent asthmagens in vitro.
Type:
text; Electronic Thesis
Degree Name:
B.S.
Degree Level:
bachelors
Degree Program:
Honors College; Microbiology
Degree Grantor:
University of Arizona
Advisor:
Boitano, Scott

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titleMeasuring A. alternata Protease Activity and Their Effects on Asthmaen_US
dc.creatorGraham, Lauren Victoriaen_US
dc.contributor.authorGraham, Lauren Victoriaen_US
dc.contributor.authorFlynn, Andrea N.en_US
dc.contributor.authorSherwood, Cara L.en_US
dc.contributor.authorSchultz, Stephanie M.en_US
dc.contributor.authorHoffman, Justinen_US
dc.contributor.authorDaines, Michael O.en_US
dc.contributor.authorBoitano, Scotten_US
dc.date.issued2013-
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractAlternaria alternata, is a key factor in childhood allergic asthma in semi-arid climates. We have shown that filtrates from A. alternata can both cause asthma symptoms in a mouse model as well as directly activate human bronchial epithelial cells, which provide the first point of contact for microbes in the airway lumen. These interactions are dependent on A. alternata protease activity within the host acting on protease activated receptor-2 (PAR₂) (1). Proteolysis of PAR₂ results in an exposed “tethered ligand” that initiates host intracellular signaling pathways and subsequent physiological response (1). In this report, we measured the protease activity of various A. alternata filtrate samples using a standard, casein-based fluorescent protease detection assay and a novel fluorescent probe we designed to include the specific proteolytic sequence of PAR₂. Both assays showed protease activity of A. alternata filtrates that matched their respective signaling responses in host epithelial cells. Conditions that limited protease activity effectively blocked protease detections in the assays and cell signaling responses. We conclude that A. alternata filtrates induce airway epithelial cell signaling and asthmatic response partly via protease activity on PAR₂. By utilizing our novel proteolytic assay we may better predict potency of various PAR₂-dependent asthmagens in vitro.en_US
dc.typetexten_US
dc.typeElectronic Thesisen_US
thesis.degree.nameB.S.en_US
thesis.degree.levelbachelorsen_US
thesis.degree.disciplineHonors Collegeen_US
thesis.degree.disciplineMicrobiologyen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorBoitano, Scott-
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