Assessing the efficacy of vesicle fusion with planar membrane arrays using a mitochondrial porin as reporter

Kamila Justyna Pszon-Bartosz, Jesper S. Hansen, Karin B. Stibius, Jesper S. Groth, Jenny Emnéus, Oliver Geschke, Claus Helix Nielsen

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Reconstitution of functionally active membrane protein into artificially made lipid bilayers is a challenge that must be overcome to create a membrane-based biomimetic sensor and separation device. In this study we address the efficacy of proteoliposome fusion with planar membrane arrays. We establish a protein incorporation efficacy assay using the major non-specific porin of Fusobacterium nucleatum (FomA) as reporter. We use electrical conductance measurements and fluorescence microscopy to characterize proteoliposome fusion with an array of planar membranes. We show that protein reconstitution in biomimetic membrane arrays may be quantified using the developed FomA assay. Specifically, we show that FomA vesicles are inherently fusigenic. Optimal FomA incorporation is obtained with a proteoliposome lipid-to-protein molar ratio (LPR)=50 more than 105 FomA proteins could be incorporated in a bilayer array with a total membrane area of 2mm2 within 20min. This novel assay for quantifying protein delivery into lipid bilayers may be a useful tool in developing biomimetic membrane applications.
Original languageEnglish
JournalBiochemical and Biophysical Research Communications
Volume406
Issue number1
Pages (from-to)96-100
ISSN0006-291X
DOIs
Publication statusPublished - 2011

Keywords

  • Fusion
  • Membranes
  • Outer membrane protein
  • Vesicles

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