Keywords

inflammation; NOD2; RIP2; XIAP; ALOX5; FLAP

Abstract

Inflammation is an essential immune response. However, dysregulated inflammation can result in chronic inflammatory diseases, like Crohn’s disease, Blau syndrome, and early-onset sarcoidosis. Receptor-interacting serine/threonine protein kinase 2 (RIP2) is a kinase that plays a critical role in nucleotide-binding oligomerization domain-containing protein 1 and 2 (NOD1 and 2) signaling pathways, ultimately triggering NF-kB activation and the secretion of pro- inflammatory cytokines. Our laboratory discovered that RIP2 also promotes the activation of arachidonate 5-Lipoxygenase (ALOX5 or 5LO), an enzyme important for producing lipid mediators. Interaction of RIP2 with the X-linked inhibitor of apoptosis protein (XIAP) has been demonstrated to be crucial for NF-kB activation and cytokine production. The purpose of this project is to determine if and how RIP2:XIAP interactions influence ALOX5 activity. This project utilizes transient transfection, competitive inhibition, fractionation, co-immunoprecipitation, SDS- PAGE, and Western blotting to analyze how disrupting RIP2:XIAP interactions influences ALOX5-activating phosphorylation and ALOX5 association with FLAP (5-Lipoxygenase Activating Protein). This study also discusses the development of a fluorescenct imaging assay to capture ALOX5:FLAP interactions. Our results indicate that RIP2:XIAP interactions are dispensable for RIP2-mediated ALOX5 activation. Understanding the mechanisms by which RIP2 influences ALOX5 will be helpful in the design, development, and testing of drugs treating NOD2-associated inflammatory diseases.

Thesis Completion Year

2025

Thesis Completion Semester

Spring

Thesis Chair

Tigno-Aranjuez, Justine

College

College of Medicine

Department

Burnett School of Biomedical Sciences

Thesis Discipline

Biomedical Sciences

Language

English

Access Status

Campus Access

Length of Campus Access

3 years

Subjects

Lipoxygenases; Inflammation--Molecular aspects; Cytokines--Research; Apoptosis--Research; Inflammation--Pathogenesis

Download

Restricted to the UCF community until 5-15-2028; it will then be open access.

Share

COinS
 

Accessibility Statement

This item was created or digitized prior to April 24, 2027, or is a reproduction of legacy media created before that date. It is preserved in its original, unmodified state specifically for research, reference, or historical recordkeeping. In accordance with the ADA Title II Final Rule, the University Libraries provides accessible versions of archival materials upon request. To request an accommodation for this item, please submit an accessibility request form.

Rights Statement

In Copyright