Porin pores of mitochondrial outer membranes from high and low eukaryotic cells: biochemical and biophysical characterization.
The mitochondrial porins from mammalian tissues and from low eukaryotic cells were purified with a high yield, and their biochemical and functional properties were investigated. When analyzed by SDS gel electrophoresis, all mammalian porins show a very similar apparent molecular mass (35-35.5 kDa). In contrast yeast and Paramecium porins have a molecular mass of 30 and 37 kDa, respectively. The peptide maps of mammalian porins are very similar although small differences are apparent between porins of different tissues of the same organism and also between those of the same tissue of different organisms. The peptide patterns of porins from yeast and Paramecium are completely different from those of mammalian porins. Antibodies raised against the rat liver porin cross-react with all the other mammalian porins but not with that of yeast. The incorporation of porins into artificial lipid bilayer membranes showed that they are able to form pores with approximately the same specific activity. The single-channel conductance is for all porins, except for that of Paramecium, about 4 nS in 1 M KCl, corresponding to an effective pore diameter of 1.7 nm. They are voltage-dependent and switch to substates at transmembrane potentials higher than 10 mV. The number of gating charges varies, however, for pores from different tissues, indicating a different sensitivity to the potential as a result of a possible different function.
De Pinto
V
V
Department of Pharmaco-Biology, University of Bari, Italy.
Ludwig
O
O
Krause
J
J
Benz
R
R
Palmieri
F
F
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D008970:Molecular Weight; D010449:Peptide Mapping; D018272:Porins; D011817:Rabbits; D051381:Rats; D013045:Species Specificity; D050994:Voltage-Dependent Anion Channels
D008565:Membrane Proteins; D018272:Porins; D050994:Voltage-Dependent Anion Channels; D000255:Adenosine Triphosphate