Quantitative Nano-Platforms for Interrogation of Curvature Sensitive Proteins

Abstract

Membrane curvature is a defining parameter which mediates many critical cellular processes like endocytosis and exocytosis. This parameter is mainly mediated by several types of curvature-sensing domains, including N-BAR, F-BAR, I-BAR, and ENTH domain. Previous studies of these curvature-sensing domains lacking precise quantification of curvature sensitivity while maintaining the ease of use the assay. Herein, we design a concise platform to quantitatively determine the curvature-sensing range for several curvature-sensing proteins, which implies the role of these proteins in several membrane-associated processes. The core of our technology consists of simple nanofabrication and microfluidics, integrated with quick lipid bilayer engineering and protein lysate preparation. This design provides us with the flexibility to switch lipid compositions, manipulate lysate components, and engineer complex curvatures, which serves as a powerful tool to investigate curvature-dependent cellular processes.