<br/>The natural mechanisms that direct proteins to membranes are typically complex, requiring multiple steps and accessory components. Because of this, it can be difficult to engineer artificial systems that allow for controlled protein localization. As a solution to this problem, we developed a modular method for membrane localization of proteins by using chemically modified phospholipid anchors capable of covalent attachment to O6-methylguanine DNA methyltransferase (SNAP-tag) fusion proteins. Our system is compatible with cell-free expression, allowing for genetic targeting of proteins to lipid membranes within giant unilamellar vesicles. We have also demonstrated that photocaged lipid precursors enable light-triggered spatial and temporal control over protein localization within populations of vesicles as well as individual vesicle membranes. Using this technique, it is possible to control protein induced membrane curvature effects, similar to what has been previously observed with certain natural membrane-bound proteins. This work addresses a current need in synthetic biology for simplified and robust methods to control membrane localization of expressed proteins and shows promise as a general tool for protein targeting to lipid vesicles and cellular membranes.

Lipid anchors for controlled localization of protein to phospholipid membranes

251st National Meeting (2016)

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