Identification of latent procathepsins B and L in microsomal lumen: Characterization of enzymatic activation and proteolytic processing in vitro

Yukio Nishimura, Takahiro Kawabata, Keitaro Kato

研究成果: ジャーナルへの寄稿記事

147 引用 (Scopus)

抄録

Procathepsins B and L in the hepatic endoplasmic lumen were identified as having a molecular weight of 39,000 by immunoblot analysis. The proenzymes were then purified to remove the mature enzymes by concanavalin A-Sepharose chromatography. The concanavalin A-adsorbed fractions containing the proenzymes showed no appreciable activities of cathepsins B and L. When those fractions were incubated at pH 3.0, the enzymatic activities markedly increased: the activities of cathepsins B and L after 36 h incubation were 60 and 210 times those of the controls, respectively. Immunoblot analysis showed that after 36 h incubation the proenzymes disappeared and the mature enzymes increased. Thus the proenzymes were processed to the mature enzymes under acidic conditions of pH 3.O. The marked increases of enzymatic activities and the conversion of the proenzymes to the mature forms were completely blocked with pepstatin, which is a potent inhibitor of aspartic proteases. The results strongly suggested that a processing protease for procathepsins B and L might be cathepsin D, a major lysosomal aspartic protease. Indeed, lysosomal cathepsin D could convert microsomal procathepsin B to the mature enzyme in vitro. Therefore, procathepsins B and L seem first to be synthesized as enzymatically inactive forms in endoplasmic reticulum and successively may be converted into active forms by cathepsin D in lysosomal compartments.

元の言語英語
ページ(範囲)64-71
ページ数8
ジャーナルArchives of Biochemistry and Biophysics
261
発行部数1
DOI
出版物ステータス出版済み - 2 15 1988

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Enzyme Precursors
Chemical activation
Cathepsin D
Cathepsin L
Processing
Cathepsin B
Peptide Hydrolases
Enzymes
Agarose Chromatography
Concanavalin A
Chromatography
Protease Inhibitors
Endoplasmic Reticulum
Molecular Weight
Molecular weight
procathepsin B
procathepsin L
In Vitro Techniques
Liver

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology

これを引用

Identification of latent procathepsins B and L in microsomal lumen : Characterization of enzymatic activation and proteolytic processing in vitro. / Nishimura, Yukio; Kawabata, Takahiro; Kato, Keitaro.

:: Archives of Biochemistry and Biophysics, 巻 261, 番号 1, 15.02.1988, p. 64-71.

研究成果: ジャーナルへの寄稿記事

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