Abstract
| Original language | English |
|---|---|
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 406 |
| Issue number | 1 |
| Pages (from-to) | 96-100 |
| ISSN | 0006-291X |
| DOIs | |
| Publication status | Published - 2011 |
Keywords
- Fusion
- Membranes
- Outer membrane protein
- Vesicles
Cite this
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Assessing the efficacy of vesicle fusion with planar membrane arrays using a mitochondrial porin as reporter. / Pszon-Bartosz, Kamila Justyna; Hansen, Jesper S.; Stibius, Karin B.; Groth, Jesper S.; Emnéus, Jenny; Geschke, Oliver; Helix Nielsen, Claus.
In: Biochemical and Biophysical Research Communications, Vol. 406, No. 1, 2011, p. 96-100.Research output: Contribution to journal › Journal article › Research › peer-review
TY - JOUR
T1 - Assessing the efficacy of vesicle fusion with planar membrane arrays using a mitochondrial porin as reporter
AU - Pszon-Bartosz, Kamila Justyna
AU - Hansen, Jesper S.
AU - Stibius, Karin B.
AU - Groth, Jesper S.
AU - Emnéus, Jenny
AU - Geschke, Oliver
AU - Helix Nielsen, Claus
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
KW - Fusion
KW - Membranes
KW - Outer membrane protein
KW - Vesicles
U2 - 10.1016/j.bbrc.2011.02.001
DO - 10.1016/j.bbrc.2011.02.001
M3 - Journal article
VL - 406
SP - 96
EP - 100
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -