Kallol Gupta, Nanobiology Institute, Yale University, and Department of Cell Biology, Yale School of Medicine

A nanoscale spatially resolved mass spectrometry-based platform for macromolecular cartography of the cellular membrane

 Kallol Gupta

¹Nanobiology Institute, Yale University, West Haven, Connecticut

²Department of Cell Biology, Yale School of Medicine, New Haven, Connecticut

ABSTRACT: The local molecular environment of the native membrane profoundly influences all aspects of membrane protein (MP) biology. Despite this, the most prevalent method of studying MPs uses detergent-like molecules that remove this critical local membrane context. This impedes our ability to quantitatively decipher the local molecular context and how it regulates structure, function, and biogenesis of MPs. Addressing this, combining a membrane-active-copolymers (MAP) library with MS based quantitative proteomics, lipidomics, and native mass spectrometry, we present a quantitative molecular platform to determine macromolecular organization of MPs and lipids in the cellular membrane with nanoscale spatial resolution. We will present key technological advancements in native mass spectrometry that enables detection and ID of signaling MP-lipid complexes directly from both in vitro and cellular membranes through electron capture based top-down fragmentation. We accompany this nativeMS-based advancements with key chemical biology tool developments that provides a high throughout and broadly applicable platform to capture the local membrane ‘nano-domains’ of a target MP directly from its endogenous organellar membrane. Interfacing such capabilities with a range of bioanalytical approaches, which include MS-based OMICS, single-molecule microscopy, EM-imaging, and biochemical assays, we will present a novel experimental avenue to render a quantitative molecular view of how the local membrane context regulates the function of MPs in human health and disease.