A water-soluble form of porin from the mitochondrial outer membrane of Neurospora crassa. Properties and relationship to the biosynthetic precursor form.
Mitochondrial porin, the outer membrane pore-forming protein, was isolated in the presence of detergents and converted into a water-soluble form. This water-soluble porin existed under nondenaturing conditions as a mixture of dimers and oligomers. The proportion of dimers increased with decreasing porin concentration during conversion. Water-soluble porin inserted spontaneously into artificial bilayers as did detergent-solubilized porin. Whereas the latter form had no specific requirements for the lipid composition of the bilayer, water-soluble porin inserted only into membranes containing a sterol, and only in the presence of very low concentrations of Triton X-100 (0.001% w/v) in the solution bathing the bilayer. The channels formed by water-soluble porin were indistinguishable from those formed by detergent-purified porin with respect to specific conductance and voltage dependence of conductance. Water-soluble porin bound tightly in a saturable fashion to isolated mitochondria. The bound form was readily accessible to added protease, indicating its presence on the mitochondrial surface. The number of binding sites was in the range of 5-10 pmol/mg of mitochondrial protein. Water-soluble porin apparently binds to a site on the assembly pathway of the porin precursor, since mitochondria whose binding sites were saturated with the water-soluble form did not import porin precursor synthesized in a cell-free system.
Pfaller
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Freitag
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Harmey
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MA
Benz
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Neupert
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D001425:Bacterial Outer Membrane Proteins / Q000032:analysis*; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D007425:Intracellular Membranes / Q000032:analysis; D008928:Mitochondria / Q000032:analysis; D008970:Molecular Weight; D009491:Neurospora / Q000032:analysis*; D009492:Neurospora crassa / Q000032:analysis* / Q000648:ultrastructure; D018272:Porins; D011498:Protein Precursors / Q000032:analysis*; D012995:Solubility; D013388:Succinimides / Q000494:pharmacology; D014867:Water
D001425:Bacterial Outer Membrane Proteins; D018272:Porins; D011498:Protein Precursors; D013388:Succinimides; D014867:Water; C019358:disuccinimidyl suberate