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J. Biol. Chem., Vol. 263, Issue 7, 3188-3193, 03, 1988
H Ono and S Tuboi
The mechanism of import of proteins into mitochondria was studied by using
the peptide of the presequence of ornithine aminotransferase (the
extrapeptide), which was chemically synthesized and is composed of 34 amino
acids. When the extrapeptide was incubated with isolated mitochondria in
the presence of a rabbit reticulocyte lysate at 25 degrees C, it was
imported into the mitochondrial matrix, and the import depended on the
inner membrane potential, but not added ATP. The import of several
precursors of mitochondrial proteins was competitively inhibited by the
presence of excess extrapeptide in the reaction system, indicating that the
extrapeptide and mitochondrial proteins were imported by the same
machinery. Import of the extrapeptide was significantly stimulated by
addition of a rabbit reticulocyte lysate, and a component of the lysate
(the cytosolic factor) stimulating import of the extrapeptide was purified
about 20,000 times by successive column chromatography on DEAE-cellulose
and aminopentyl-Sepharose 4B. The binding of the extrapeptide to liposomes
composed of egg lecithin and partially purified receptor of the precursor
of mitochondrial protein (Ono, H., and Tuboi, S., (1985) Biochem. Int. 10,
351-357) required the cytosolic factor when the concentration of the
peptide was less than 1.5 X 10(-8) M, suggesting that the physiological
binding of the precursors of mitochondrial proteins to the receptor is
dependent on the cytosolic factor. The extrapeptide and the cytosolic
factor were shown to form a complex. From these results, the mechanism of
binding of the extrapeptide to the receptor of the mitochondrial outer
membrane is suggested to be as follows: the peptide (the precursor of
mitochondrial protein) and the cytosolic factor form a complex, and then
the complex is recognized by and bound to the receptor.
The cytosolic factor required for import of precursors of mitochondrial proteins into mitochondria
Department of Biochemistry, Yamagata University School of Medicine, Japan.
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