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The Structure and Assembly of Membrane Proteins in Native Membranes studied by AFM (MEM-STRUCT-AFM)
Date du début: 1 janv. 2013, Date de fin: 31 déc. 2017 PROJET  TERMINÉ 

One of the key characteristics of life is the existence of boundaries that delineate the organism from its environment. The boundary membrane structure that defines all cells is called the plasma membrane. Membranes are constituted of a large variety of lipids and membrane proteins, key molecules for plenty vital cellular functions such as transport, energy transduction, signaling, and communication, to name a few. Despite the importance of membrane processes, little is known about the structure of its constituents and less about their supramolecular arrangement. In this project the focus is set on the analysis of membrane proteins, their supramolecular complexes, their structural assembly and mechanism of cooperative function, and thus an integral view of the native membrane.To achieve this goal, we are performing membrane biochemistry and develop 3 axes of research, all Atomic Force Microscopy (AFM) –based, comprising technical developments of high-resolution and high-speed AFM, AFM-hybrid techniques, but also target-focused application research, all, to gain information about the structure and assembly of membrane proteins in native membranes at unprecedented resolution.1) Technical developments: In this project subaxis, we are performing two major technical developments, a hybrid atomic force microscope (AFM) - tip enhanced raman spectroscopy (AFM-TERS), and a high-speed AFM - Optical Microscopy setup.2) AFM imaging: Analysis of the supramolecular assembly of membrane complexes in native membranes ex cellula and in cellula, with a particular focus on alterations of membrane structure and membrane protein assembly and conformation due to pathology or addition of drugs.3) AFM force spectroscopy: Analysis of forces and affinities within and in between membrane complexes on molecules and cells, with a particular focus on the development of small-cantilever HS-AFM -based force spectroscopy (dynamic force spectroscopy and force clamp modes).