Characterization of adrenodoxin precursor expressed in Escherichia coli

Jun Iwahashi, Shigeki Furuya, Katsuyoshi Mihara, Tsuneo Omura

研究成果: ジャーナルへの寄稿記事

23 引用 (Scopus)

抄録

The precursor of bovine adrenodoxin (pAd), a mitochondrial protein, was expressed in Escherichia coil. The cloned cDNA of pAd was ligated to an expression vector pET-3d, and silent mutations were introduced into the N-terminal portion of the cDNA in order to increase the expression. The precursor was highly expressed (approximately 20% of the total cell protein) as the inclusion body, and contained an iron-sulfur center as judged from its optical absorption spectra. The inclusion body was solubilized with 7 M urea and pAd was purified in the presence of urea. The purified pAd was efficiently imported into isolated bovine adrenal cortex mitochondria and processed to the mature form. The import reaction required ATP inside the mitochondria in addition to the inner membrane potential, and was strongly inhibited by trypsin treatment of the mitochondria, as in the case of the in vitro translated precursor. It was, however, not dependent on the unfolding activity of the cytosolic factor with extramitochondrial ATP.

元の言語英語
ページ(範囲)451-455
ページ数5
ジャーナルJournal of biochemistry
111
発行部数4
DOI
出版物ステータス出版済み - 1 1 1992

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Adrenodoxin
Escherichia coli
Mitochondria
Inclusion Bodies
Urea
Complementary DNA
Adenosine Triphosphate
Mitochondrial Proteins
Escherichia
Sulfur
Trypsin
Light absorption
Adrenal Cortex
Absorption spectra
Iron
Membrane Potentials
Membranes
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

これを引用

Characterization of adrenodoxin precursor expressed in Escherichia coli. / Iwahashi, Jun; Furuya, Shigeki; Mihara, Katsuyoshi; Omura, Tsuneo.

:: Journal of biochemistry, 巻 111, 番号 4, 01.01.1992, p. 451-455.

研究成果: ジャーナルへの寄稿記事

Iwahashi, Jun ; Furuya, Shigeki ; Mihara, Katsuyoshi ; Omura, Tsuneo. / Characterization of adrenodoxin precursor expressed in Escherichia coli. :: Journal of biochemistry. 1992 ; 巻 111, 番号 4. pp. 451-455.
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