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1.
Biofizika ; 55(5): 803-8, 2010.
Article in Russian | MEDLINE | ID: mdl-21033345

ABSTRACT

A low-molecular-weight component (LMC) inducing selective transport of calcium across the bilayer lipid membrane has been isolated from mitochondria of the bovine heart by the method developed in our laboratory, which excludes the use of detergents and proteolytic enzymes. It was shown that, in the presence of 10 mM CaCl2, LMC forms conduction channels in the membrane multiples of 5 pS. The specific inhibitor of mitochondrial calcium uniporter, ruthenium red, closes Ca2(+)-induced channels formed in the membrane by LMC. In the absence of calcium or in the presence of potassium ions only, the component is incapable of forming channels of conduction. It was shown using nuclear magnetic resonance that LMC is a complex consisting of lipids, amino acids, and sugars with a molecular weight of 1-2 kDa.


Subject(s)
Calcium Channels/chemistry , Mitochondrial Proteins/chemistry , Animals , Calcium/chemistry , Calcium Channel Blockers/chemistry , Calcium Channels/isolation & purification , Cations, Divalent , Cattle , Ion Transport , Lipid Bilayers/chemistry , Magnetic Resonance Spectroscopy , Mitochondrial Proteins/isolation & purification , Ruthenium Red/chemistry
2.
Biochemistry (Mosc) ; 70(2): 187-94, 2005 Feb.
Article in English | MEDLINE | ID: mdl-15807658

ABSTRACT

Calcium ions are of central importance in cellular physiology, as they carry the signal activating cells to perform their programmed function. Ca(2+) is particularly suitable for this role because of its chemical properties and because its free concentration gradient between the extra-cellular and the cytosolic concentrations is very high, about four orders of magnitude. The cytosolic concentration of Ca(2+) is regulated by binding and chelation by various substances and by transport across plasma and intracellular membranes. Various channels, transport ATPases, uniporters, and antiporters in the plasma membrane, endoplasmic and sarcoplasmic reticulum, and mitochondria are responsible for the transport of Ca(2+). The regulation of these transport systems is the subject of an increasing number of studies. In this short review, we focus on the mitochondrial transporters, i.e. the calcium uniporter used for Ca(2+) uptake, and the antiporters used for the efflux, i.e. the Ca(2+)/Na(+) antiporter in mitochondria and the plasma membrane of excitable cells, and the Ca(2+)/nH(+) antiporter in liver and some other mitochondrial types. Mitochondria are of special interest in that Ca(2+) stimulates respiration and oxidative phosphorylation to meet the energy needs of activated cells. The studies on Ca(2+) and mitochondria began in the fifties, but interest in mitochondrial Ca(2+) handling faded in the late seventies since it had become apparent that mitochondria in resting cells contain very low Ca(2+). Interest increased again in the nineties also because it was discovered that mitochondria and Ca(2+) had a central role in apoptosis and necrosis. This is of special interest in calcium overload and oxidative stress conditions, when the opening of the mitochondrial permeability transition pore is stimulated.


Subject(s)
Calcium/physiology , Mitochondria/metabolism , Animals , Calcium/metabolism , Calcium Signaling/physiology , Cell Membrane/metabolism , Endoplasmic Reticulum/metabolism
3.
Toxicol In Vitro ; 17(5-6): 737-44, 2003.
Article in English | MEDLINE | ID: mdl-14599471

ABSTRACT

The in vitro boar spermatozoon test was compared with the LC ion trap MS analysis for measuring the cereulide content of a pasta dish, implemented in serious emetic food poisoning caused by Bacillus cereus. Both assays showed that the poisonous food contained approximately 1.6 microg of cereulide g(-1) implying the toxic dose in human as < or =8 microg kg(-1) body weight. The threshold concentration of cereulide provoking visible mitochondrial damage in boar sperm exposed in vitro was 2 ng of cereulide ml(-1) of extended boar sperm. The same threshold value was found for cereulide extracted from the food and from the cultured bacteria. This shows that other constituents of the food did not enhance or mask the effects of cereulide. Exposure of four human cell lines (HeLa, Caco-2, Calu-3 and Paju) to cereulide showed that the threshold concentration for the loss of mitochondrial membrane potential in human cells was similar to that observed in boar sperm. Human cells and boar sperm were equally sensitive to cereulide. The results show that boar spermatozoan assay is useful for detecting cereulide concentrations toxic to humans. Spermatozoa in commercially available extended fresh boar and cryopreserved bull semen were compared, boar sperms were 100 times more sensitive to cereulide than bull sperms.


Subject(s)
Bacillus cereus , Bacterial Toxins/toxicity , Depsipeptides , Emetics/toxicity , Mitochondria/drug effects , Peptides, Cyclic/toxicity , Toxicity Tests/methods , Animals , Bacillus cereus/chemistry , Bacillus cereus/metabolism , Bacterial Toxins/analysis , Biomass , Caco-2 Cells/drug effects , Caco-2 Cells/pathology , Cattle , Emetics/analysis , Food Analysis , Foodborne Diseases , HeLa Cells/drug effects , HeLa Cells/pathology , Humans , Male , Membrane Potentials/drug effects , Peptides, Cyclic/analysis , Plant Extracts/poisoning , Sperm Motility/drug effects , Spermatozoa/drug effects , Spermatozoa/pathology , Swine , Triticum/chemistry
4.
FEBS Lett ; 7(4): 311-313, 1970 May 01.
Article in English | MEDLINE | ID: mdl-11947501
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