Biosynthesis and Processing of Lysosomal Cathepsin D in Primary Cultures of Rat Hepatocytes

Yukio Nishimura, Keitaro Kato, Masaru Himeno, Furuno Koji

Research output: Contribution to journalArticle

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Abstract

To investigate the intracellular transport and maturation of lysosomal cathepsin D, we carried out an in vivo pulse-chase analysis with [35S] methionine in the primary cultures of rat hepatocytes. Cathepsin D was initially synthesized as a proenzyme of 45 kDa. The proenzyme was subsequently processed, becoming a mature enzyme of 43 kDa. The proenzyme and mature enzyme showed complete susceptibility to endoglycosidase H treatment, suggesting the presence of high-mannose type oligosaccharide chains. The effects of tunicamycin and chloroquine were also investigated. In the presence of tunicamycin, the 42.5-kDa unglycosylated precursor polypeptide appeared in the cell, and this protein was exclusively secreted from the cells without undergoing proteolytic processing. These results support the notion that the oligosaccharide moieties are of importance in addressing the lysosomal hydrolases to the lysosomes. However, in the presence of chloroquine, proteolytic processing of the proenzyme was prevented, and the enhanced release of proenzyme from the cells was observed. These results indicate that the processing of proenzyme to mature enzyme would take place in the lysosomes.

Original languageEnglish
Pages (from-to)825-828
Number of pages4
JournalBiological and Pharmaceutical Bulletin
Volume18
Issue number6
DOIs
Publication statusPublished - Jan 1 1995

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Cathepsin D
Enzyme Precursors
Hepatocytes
Tunicamycin
Chloroquine
Lysosomes
Oligosaccharides
Enzymes
Glycoside Hydrolases
Hydrolases
Mannose
Methionine
Peptides
Proteins

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Pharmaceutical Science

Cite this

Biosynthesis and Processing of Lysosomal Cathepsin D in Primary Cultures of Rat Hepatocytes. / Nishimura, Yukio; Kato, Keitaro; Himeno, Masaru; Koji, Furuno.

In: Biological and Pharmaceutical Bulletin, Vol. 18, No. 6, 01.01.1995, p. 825-828.

Research output: Contribution to journalArticle

Nishimura, Yukio ; Kato, Keitaro ; Himeno, Masaru ; Koji, Furuno. / Biosynthesis and Processing of Lysosomal Cathepsin D in Primary Cultures of Rat Hepatocytes. In: Biological and Pharmaceutical Bulletin. 1995 ; Vol. 18, No. 6. pp. 825-828.
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