Cholesterol- and sphingolipid-enriched membrane nanodomains known as membrane rafts are widely recognized for their role in regulating membrane protein trafficking and function. The movement of specific proteins into or out of rafts has also been linked to a number of diseases. Despite this, few methods to experimentally perturb rafts currently exist, and we still lack a clear understanding of how rafts control cellular functions. To address these long-standing questions, we are utilizing giant plasma membrane vesicles (GPMVs) as a model to investigate mechanisms controlling raft structure, composition, and function. Isolated from the plasma membrane of live cells in the form of cell-sized vesicles, GPMVs maintain the compositional complexity of cell membranes, yet are capable of forming coexisting raft-like and non-raft phases that can be readily visualized by fluorescence microscopy. Here, I will describe new computational approaches developed in our group to facilitate high content imaging of rafts in GPMVs and discuss how we are using these methods to perform unbiased high throughput screens of small molecule libraries to discover new pharmacological approaches to manipulate raft structure and function. Ultimately, these studies should lead to the development of new classes of raft modulators that can be used to address the many still-unanswered questions about these poorly understood structures in biological membranes.

High content imaging to identify novel pharmacological modulators of membrane rafts

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.