Simultaneous measurement of ionic current and fluorescence from single protein pores.
The ability to simultaneously monitor both the ionic current and fluorescence from membrane channels and pores has the potential to link structural changes with function in such proteins. We present a new method for simultaneously measuring single-channel electrical currents and fluorescence from membrane proteins by using water-in-oil droplet bilayers. We demonstrate the simultaneous fluorescence and electrical detection of stochastic blocking by cyclodextrin in multiple staphylococcal alpha-hemolysin pores. The combined fluorescence signal from individual pores exhibits the same sequence of blocking events as the total current recording, showing that the two signals from each pore are correlated.
Heron
Andrew J
AJ
Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, UK.
Thompson
James R
JR
Cronin
Bríd
B
Bayley
Hagan
H
Wallace
Mark I
MI
D001427:Bacterial Toxins / Q000737:chemistry* / Q000378:metabolism; D004566:Electrodes; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000378:metabolism; D020100:Hydrogels / Q000737:chemistry; D007473:Ion Channels / Q000737:chemistry* / Q000378:metabolism; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010713:Phosphatidylcholines / Q000737:chemistry; D013269:Stochastic Processes; D047392:beta-Cyclodextrins / Q000737:chemistry* / Q000378:metabolism
D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D020100:Hydrogels; D007473:Ion Channels; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D047392:beta-Cyclodextrins; C021091:staphylococcal alpha-toxin; C000609658:heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin; C021016:1,2-diphytanoylphosphatidylcholine