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Updated: Jul 11, 2026

Layer-by-layer Collagen Deposition in Microfluidic Devices for Microtissue Stabilization
Published on: September 29, 2015
Synthesis of lipidated green fluorescent protein and its incorporation in supported lipid bilayers
Michael J Grogan1, Yoshihisa Kaizuka, Rosemary M Conrad
1Departments of Chemistry and Molecular and Cell Biology, Physical Biosciences and Materials Sciences Divisions, University of California-Berkeley, Berkeley, CA 94720, USA.
Abstract:
Herein we report a semisynthetic method of producing membrane-anchored proteins. Ligation of synthetic lipids with designed anchor structures to proteins was performed using native chemical ligation (NCL) of a C-terminal peptide thioester and an N-terminal cysteine lipid. This strategy mimics the natural glycosylphosphatidylinositol (GPI) linkage found in many natural membrane-associated proteins; however, the synthetic method utilizes simple lipid anchors without glycans. Synthetically lipidated recombinant green fluorescent protein (GFP) was shown to be stably anchored to the membrane, and its lateral fluidity was quantitatively characterized by direct fluorescence imaging in supported membranes. Circumventing the steps of purification from native cell membranes, this methodology facilitates the reconstitution of membrane-associated proteins.
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