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

研究成果: ジャーナルへの寄稿記事

82 引用 (Scopus)

抄録

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.

元の言語英語
ページ(範囲)416-425
ページ数10
ジャーナルJournal of biochemistry
116
発行部数2
DOI
出版物ステータス出版済み - 1 1 1994

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Cloning
Liver
Cytosol
14-3-3 Proteins
Protein Precursors
Rats
Organism Cloning
Complementary DNA
Mitochondrial Proteins
Triticum
Adrenodoxin
Proteins
Escherichia coli Proteins
Gene Library
Recombinant Proteins
Depolymerization
Mitochondria
Polyacrylamide Gel Electrophoresis
Adenosine Triphosphate
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

これを引用

CDN a cloning and characterization of mitochondrial import stimulation factor (MSF) purified from rat liver cytosol. / Alam, Rashidul; Hachiya, Naomi; Sakaguchi, Masao; Kawabata, Shun-Ichiro; Iwanaga, Sadaaki; Kitajima, Masato; Mihara, Katsuyoshi; Omura, Tsuneo.

:: Journal of biochemistry, 巻 116, 番号 2, 01.01.1994, p. 416-425.

研究成果: ジャーナルへの寄稿記事

Alam, Rashidul ; Hachiya, Naomi ; Sakaguchi, Masao ; Kawabata, Shun-Ichiro ; Iwanaga, Sadaaki ; Kitajima, Masato ; Mihara, Katsuyoshi ; Omura, Tsuneo. / CDN a cloning and characterization of mitochondrial import stimulation factor (MSF) purified from rat liver cytosol. :: Journal of biochemistry. 1994 ; 巻 116, 番号 2. pp. 416-425.
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abstract = "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.",
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AU - Alam, Rashidul

AU - Hachiya, Naomi

AU - Sakaguchi, Masao

AU - Kawabata, Shun-Ichiro

AU - Iwanaga, Sadaaki

AU - Kitajima, Masato

AU - Mihara, Katsuyoshi

AU - Omura, Tsuneo

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N2 - 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.

AB - 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.

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