CDN a cloning and characterization of mitochondrial import stimulation factor (MSF) purified from rat liver cytosol

Rashidul Alam, Naomi Hachiya, Masao Sakaguchi, Shun Ichiro Kawabata, Sadaaki Iwanaga, Masato Kitajima, Katsuyoshi Mihara, Tsuneo Omura

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85 Citations (Scopus)


We identified a liver cytosolic protein factor that stimulated the import of wheat germ lysate-synthesized precursor proteins into mitochondria. It was termed mitochondrial import stimulation factor or MSF [Hachiya, N. et al (1993) EMBO J. 12, 1579-1586]. It consisted of 32-kDa (MSFL) and 30-kDa (MSFS) polypeptides as assessed by SDS-PAGE. MSF recognized the presequence portion of mitochondrial precursor proteins and catalyzed the depolymerization and unfolding of in vitro synthesized mitochondrial precursor proteins in an ATP-dependent manner. We report here the cDNA cloning and characterization of MSF. Microsequencing of MSFL and MSFS showed that they belonged to a highly conserved, widely distributed eukaryotic protein family, collectively designated as 14-3-3 proteins. We cloned the cDNA of MSFL and that of one component of MSFS (MSFSl) from a rat liver cDNA library- The cloned cDNAs were separately expressed in Escherichia coli and the expressed proteins were purified to homogeneity. The purified recombinant MSFL and MSFSl stimulated mitochondrial import of adrenodoxin precursor (pAd) synthesized in vitro with wheat germ lysate translation system. Recombinant MSFL or MSFSl had the ability to bind with denatured pAd and they kept the precursor in an import-competent state. Rabbit polyclonal antibodies raised against the recombinant proteins inhibited the import-stimulation activity of rat liver cytosol as well as that of authentic purified MSF. Identification of MSF as 14-3-3 proteins establishes a novel function for this family of proteins and indicates their role as cytosolic chaperones to aid many important cellular events.

Original languageEnglish
Pages (from-to)416-425
Number of pages10
JournalJournal of biochemistry
Issue number2
Publication statusPublished - Aug 1994

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology


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