Activity-based protein profiling of rhomboid intramembrane proteases
Location
Science Center: Bent Corridor
Document Type
Poster
Start Date
4-28-2023 12:00 PM
End Date
4-28-2023 2:00 PM
Abstract
Rhomboid intramembrane proteases (RIPs) are a subclass of serine proteases responsible for cleaving polypeptide chains in other proteins. These enzymes have been associated with various metabolic, neurodegenerative, and parasitic diseases including type II diabetes and Alzhheimer’s disease. However, for many RIPs, no substrates have been reported, and our knowledge of the physiological pathways of these enzymes is still relatively limited. A valuable alternative tool to study these enzymes is activity-based protein profiling (ABPP), which utilizes small molecule probes to monitor protein activity. My current project is focusing on synthesizing a library of potential activity-based probes, including β-lactams, benzoxazinones and isocoumarins, and testing for their ability to engage bacterial (GlpG) and mammalian RIPs (RHBDL1, RHBDL2, RHBDL3, RHBDL4, and PARL). We are using the azide-alkyne Huisgen cycloaddition reaction with a functionalized rhodamine as a means to visualize labeling of these proteins. Using these methods, we have already observed probe engagement of GlpG, RHBDL2, and RHBDL3 with some of our initial structures. These promising initial results could provide a useful foundation for understanding the physiological roles of human RIPs and could inspire the development of molecules that could alter the activity of these enzymes.
Keywords:
Enzymes, Chemical probes, Membrane proteins, Chemical biology
Recommended Citation
Nguyen, Dat, "Activity-based protein profiling of rhomboid intramembrane proteases" (2023). Research Symposium. 2.
https://digitalcommons.oberlin.edu/researchsymp/2023/posters/2
Major
Biochemistry/Chemistry
Project Mentor(s)
William H. Parsons, Geoscience
2023
Activity-based protein profiling of rhomboid intramembrane proteases
Science Center: Bent Corridor
Rhomboid intramembrane proteases (RIPs) are a subclass of serine proteases responsible for cleaving polypeptide chains in other proteins. These enzymes have been associated with various metabolic, neurodegenerative, and parasitic diseases including type II diabetes and Alzhheimer’s disease. However, for many RIPs, no substrates have been reported, and our knowledge of the physiological pathways of these enzymes is still relatively limited. A valuable alternative tool to study these enzymes is activity-based protein profiling (ABPP), which utilizes small molecule probes to monitor protein activity. My current project is focusing on synthesizing a library of potential activity-based probes, including β-lactams, benzoxazinones and isocoumarins, and testing for their ability to engage bacterial (GlpG) and mammalian RIPs (RHBDL1, RHBDL2, RHBDL3, RHBDL4, and PARL). We are using the azide-alkyne Huisgen cycloaddition reaction with a functionalized rhodamine as a means to visualize labeling of these proteins. Using these methods, we have already observed probe engagement of GlpG, RHBDL2, and RHBDL3 with some of our initial structures. These promising initial results could provide a useful foundation for understanding the physiological roles of human RIPs and could inspire the development of molecules that could alter the activity of these enzymes.
