Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 22, Issue 12, Pages -Publisher
MDPI
DOI: 10.3390/ijms22126335
Keywords
in vitro reconstitution; liposome; functionalised GUVs; adhesion
Funding
- French National Research Agency [ANR-18-CE13-0026]
- Agence Nationale de la Recherche (ANR) [ANR-18-CE13-0026] Funding Source: Agence Nationale de la Recherche (ANR)
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This study describes a new method for preparing GUVs incorporating the transmembrane protein integrin, with successful incorporation of functional integrin molecules.
Giant unilamellar vesicles (GUV) are powerful tools to explore physics and biochemistry of the cell membrane in controlled conditions. For example, GUVs were extensively used to probe cell adhesion, but often using non-physiological linkers, due to the difficulty of incorporating transmembrane adhesion proteins into model membranes. Here we describe a new protocol for making GUVs incorporating the transmembrane protein integrin using gel-assisted swelling. We report an optimised protocol, enumerating the pitfalls encountered and precautions to be taken to maintain the robustness of the protocol. We characterise intermediate steps of small proteoliposome formation and the final formed GUVs. We show that the integrin molecules are successfully incorporated and are functional.
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