Nanopore-spanning proteolipid membranes
Abstract
Functional investigations of membrane proteins require probing them in their native environment: the lipid bilayer. The presence of a lipid bilayer is required to ensure correct folding and function. Due to the fragility of membrane protein conformation, this tends to require either extremely complicated and case-by-case developed detergent assays to extract and reconstitute membrane proteins or the direct use of native membranes. Today, the majority of functional assays, e.g. for drug screening, are carried out using fluorescently labeled ligands directly on live cells; this limits investigations to affinity assays and with all the well-known drawbacks of false positives and non-quantitative response of fluorescence labels and cell systems. These limitations could be a reason for the increasingly severe failure of pharma to deliver new drugs, and is as well a strong driver of costs. I propose to explore recent advances from my institute and my own research to implement a surface-based membrane platform that could allow investigating membrane proteins reconstituted in liposomes or even directly from native liposomes extracted from cells, on a nanoplasmonic chip that also could allow functional studies and to do so without denaturing the membrane protein at the surface. The nanostructured and nanoplasmonic chip is being developed as a side-track in my current project, and recent results have led to the development of the hypotheses put forward in this proposal regarding improved methods to integrate transmembrane proteins into supported membrane sensor platforms. Using nanostructured chips I will in this project explore two promising and key technological steps: 1. The use of PEG-lipid and PEG-liposomes to facilitate rupture of native liposomes without denaturing membrane protein components and keep them functional and mobile in surface-based assays. 2. The selective fusion of membrane-protein containing vesicles over nanopores containing (nanoplasmonic and electrochemical) sensor elements. A completed project will result in increased understanding of interaction of liposomes with nanostructures and substrates functionalized on the nanoscale, i.e. nanoscale sensors. It will provide new tools for insertion of membrane proteins into surface-based sensor structures functionalized with membranes.
keywords lipid biosensor self-assembly nanostructure Artificial membrane
Publikationen
Project staff
Erik Reimhult
Univ.Prof. Dr. Erik Reimhult
erik.reimhult@boku.ac.at
Tel: +43 1 47654-80211
BOKU Project Leader
01.03.2017 - 31.08.2018
BOKU partners
External partners
AIT Austrian Institute of Technology GmbH
Nicolaus Leitner
partner