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. 2007 Apr 25;12(4):493–508. doi: 10.2478/s11658-007-0019-9

Stilbene derivatives inhibit the activity of the inner mitochondrial membrane chloride channels

Izabela Koszela-Piotrowska 1,✉, Katarzyna Choma 1,2, Piotr Bednarczyk 1,2, Krzysztof Dołowy 2, Adam Szewczyk 1, Wolfram S Kunz 3, Lubica Malekova 4, Viera Kominkova 4, Karol Ondrias 4
PMCID: PMC6275615  PMID: 17457523

Abstract

Ion channels selective for chloride ions are present in all biological membranes, where they regulate the cell volume or membrane potential. Various chloride channels from mitochondrial membranes have been described in recent years. The aim of our study was to characterize the effect of stilbene derivatives on single-chloride channel activity in the inner mitochondrial membrane. The measurements were performed after the reconstitution into a planar lipid bilayer of the inner mitochondrial membranes from rat skeletal muscle (SMM), rat brain (BM) and heart (HM) mitochondria. After incorporation in a symmetric 450/450 mM KCl solution (cis/trans), the chloride channels were recorded with a mean conductance of 155 ± 5 pS (rat skeletal muscle) and 120 ± 16 pS (rat brain). The conductances of the chloride channels from the rat heart mitochondria in 250/50 mM KCl (cis/trans) gradient solutions were within the 70–130 pS range. The chloride channels were inhibited by these two stilbene derivatives: 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS) and 4-acetamido-4′-isothiocyanostilbene-2,2′-disulfonic acid (SITS). The skeletal muscle mitochondrial chloride channel was blocked after the addition of 1 mM DIDS or SITS, whereas the brain mitochondrial channel was blocked by 300 μM DIDS or SITS. The chloride channel from the rat heart mitochondria was inhibited by 50–100 μM DIDS. The inhibitory effect of DIDS was irreversible. Our results confirm the presence of chloride channels sensitive to stilbene derivatives in the inner mitochondrial membrane from rat skeletal muscle, brain and heart cells.

Keywords: Mitochondria, Chloride channel, Stilbene derivatives, Black lipid membrane

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Abbreviations used

BLM

black lipid membrane

BM

brain mitochondria

CLICs

chloride intracellular channels

DIDS

4,4′-diisothiocyanostilbene-2,2′-disulfonic acid

DTNB

6,6′-dinitro-3,3′-dithiodibenzoic acid

HM

heart mitochondria

ITS

4-acetamido-4-isothiocyanostilbene-2,2′-disulfonic acid

IMAC

mitochondrial inner membrane anion channel

SITS

4-acetamido-4-isothiocyanostilbene-2,2′-disulfonic acid

SMM

skeletal muscle mitochondria

SMP

submitochondrial particles

TNFα

tumor necrosis factor α

UPCs

uncoupling proteins

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