Thylakoid membranes contain a high-conductance channel.
Ion channels in the thylakoid membrane were investigated by direct patch clamping on swollen thylakoids. A preparation method has been developed in order to release osmotically swollen intact thylakoids from pea protoplasts derived from cotyledons of young Pisum sativum plants. The swollen thylakoids with typical diameters between 10 microm and 20 microm formed reproducibly high-resistance seals with patch pipettes. We observed a potassium channel with a main conductant state of lambda approximately 40 pS and a conductance of lambda approximately 90 pS (in asymmetric 20/100 mM KCl) for the fully open channel. Surprisingly, the thylakoid membranes also contained a high-conductance channel with a main conductant state of lambda approximately 620 pS (in asymmetric 20/100 mM KCl), revealing also higher and lower conductant states. With a different experimental approach we showed that thylakoids are able to accumulate transiently the membrane impermeant fluorescent dye Lucifer Yellow which likewise suggests the presence of a pore-like channel with a diameter large enough to allow permeation of Lucifer Yellow.
Hinnah
S C
SC
Biophysik, Universität Osnabrück, FB Biologie/Chemie, Germany.
Wagner
R
R
D005456:Fluorescent Dyes; D007425:Intracellular Membranes / Q000502:physiology*; D015640:Ion Channel Gating; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology*; D015636:Magnesium Chloride / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018715:Microscopy, Video; D015388:Organelles / Q000502:physiology*; D018408:Patch-Clamp Techniques; D018532:Pisum sativum / Q000502:physiology*; D015221:Potassium Channels / Q000494:pharmacology; D011523:Protoplasts / Q000502:physiology; D013997:Time Factors
D005456:Fluorescent Dyes; D007473:Ion Channels; D015221:Potassium Channels; D015636:Magnesium Chloride