cDNA Cloning and Characterization of Mitochondrial Import Stimulation Factor (MSF) Purified from Rat Liver Cytosol.

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概要

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• 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 (MSFS1) 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 MSFS1 stimulated mitochondrial import of adrenodoxin precursor (pAd) synthesized in vitro with wheat germ lysate translation system. Recombinant MSFL or MSFS1 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.

• 社団法人 日本生化学会の論文

著者

• Omura Tsuneo

  Department Of Biochemistry Vanderbilt University School Of Medicine

• Kawabata Shun-ichiro

  Department Of Bilolgy Faculty Of Science Kyushu University

• Sakaguchi Masao

  Department Of Electronics And Control Engineering Nagano National College Of Technology

• Iwanaga Sadaaki

  Department Of Biology Faculty Of Scienc Kyushu University

• Mihara Katsuyoshi

  Department Of Molecular Biology Graduate School Of Medical Science

• Hachiya Naomi

  Department Of Cortical Function Disorders National Institute Of Neuroscience National Center Of Neur

• Kawabata Shun-ichiro

  Department of Molecular Biology, Graduate School of Medical Science, Kyushu University

• Alam Rashidul

  Department of Molecular Biology, Graduate School of Medical Science, Kyushu University

• Kitajima Masato

  Computer Chemistry Systems Department, Fujitsu Limited

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