Asymmetrical labeling of giant phospholipid vesicles.
The method for labeling of inner membrane leaflet in unilamellar giant POPC vesicles was developed and characterised. Symmetrically NBD-PC labeled vesicles were treated by sodium dithionite, which undergoes an irreversible chemical reaction with NBD-PC molecule making it non-fluorescent. After the addition of dithionite the fluorescence on single vesicles as well as on vesicle suspension showed a 50% decrease of its initial value corresponding to marker quenching in the outer leaflet. Hence, fluorimetry as well as fluorescence microscopy prove that dithionite quenching is a suitable method to induce an asymmetrical labeling of the NBD-PC marked giant vesicles.
Gomiscek
G
G
Institute of Biophysics, Faculty of Medicine, University of Ljubljana, Slovenia.
Arrigler
V
V
Gros
M
M
Yupncic
M
M
Svetina
S
S
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives*; D004227:Dithionite / Q000494:pharmacology; D005470:Fluorometry; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence; D010713:Phosphatidylcholines; D010743:Phospholipids / Q000378:metabolism*
C078632:1-acyl-2-(12-((7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)dodecanoyl)phosphatidylcholine; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D010743:Phospholipids; D004227:Dithionite; D009327:4-Chloro-7-nitrobenzofurazan