Patch-clamp experiments with porins extracted from a marine bacterium (Photobacterium profundum strain SS9) and reconstituted in liposomes.
The reconstitution of bacterial porins in liposome bilayers for patch-clamp recording is well established. However, the solutions used in the dehydration, rehydration, and osmotic swelling of the liposomes have been developed for porins from enteric bacteria. Porins from marine bacteria normally function in contact with seawater whose ionic composition and osmotic pressure would appear to be incompatible with the established methods. Here, we show that, contrary to expectation, an established reconstitution and patch-clamp method works well with porins, mainly OmpH and OmpL, extracted from the deep-sea marine bacterium Photobacterium profundum strain SS9 and that seawater can be introduced at a supplementary stage.
Macdonald
A G
AG
Department of Biomedical Sciences, University of Aberdeen, Aberdeen AB24 2TZ, Scotland, UK. a.macdonald@abdn.ac.uk
Martinac
B
B
Bartlett
D H
DH
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001426:Bacterial Proteins; D002103:Cadaverine / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000378:metabolism*; D018408:Patch-Clamp Techniques / Q000379:methods*; D010776:Photobacterium / Q000378:metabolism*; D018272:Porins / Q000737:chemistry* / Q000378:metabolism; D011312:Pressure; D013696:Temperature; D013997:Time Factors
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008081:Liposomes; C491058:OmpL protein, E coli; C100900:OmpL protein, bacteria; D018272:Porins; C067128:ompH protein, bacteria; D002103:Cadaverine