Isolation and sequencing of a cDNA clone encoding rat liver lysosomal cathepsin D and the structure of three forms of mature enzymes

Hideaki Fujita, Yoshitaka Tanaka, Youichiro Noguchi, Akira Kono, Masaru Himeno, Keitaro Kato

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

We isolated and sequenced a cDNA clone corresponding to the entire coding sequence of rat liver lysosomal cathepsin D. The deduced amino acid sequence revealed that cathepsin D consists of 407 amino acid residues (Mr 44,608) and the 20 NH2-terminal residues seem to constitute a cleavable signal peptide after which 44 amino acid residues follow as a propeptide. Two putative N-linked glycosylation sites and aspartic acid in the active site are as well conserved as those of human lysosomal cathepsin D. In the NH2-terminal sequence analysis of two isolated heavy chains of the mature enzyme, the termini were assigned as tryptophan (118th residue) and glycine (165th or 166th residue), respectively, hence demonstrates that the two heavy chains derive from a split of the single chain of cathepsin D at position between 117th and 118th or between 164th and 165th or 165th and 166th amino acids. We conclude that cathepsin D in rat liver lysosomes is a mixture of three forms composed of a single and two two-chain forms. However, the amounts of the two two-chain forms are low compared with that of the single chain form. Densidometric determination after SDS-PAGE revealed that the two two-chain forms account for less than 5% of the single chain form. There is a 82% similarity in amino acid level between rat and human liver lysosomal cathepsin D.

Original languageEnglish
Pages (from-to)190-196
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume179
Issue number1
DOIs
Publication statusPublished - Aug 30 1991

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Cathepsin D
Liver
Rats
Complementary DNA
Clone Cells
Amino Acids
Enzymes
Glycosylation
Protein Sorting Signals
Lysosomes
Aspartic Acid
Tryptophan
Glycine
Sequence Analysis
Polyacrylamide Gel Electrophoresis
Amino Acid Sequence
Catalytic Domain

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Isolation and sequencing of a cDNA clone encoding rat liver lysosomal cathepsin D and the structure of three forms of mature enzymes. / Fujita, Hideaki; Tanaka, Yoshitaka; Noguchi, Youichiro; Kono, Akira; Himeno, Masaru; Kato, Keitaro.

In: Biochemical and Biophysical Research Communications, Vol. 179, No. 1, 30.08.1991, p. 190-196.

Research output: Contribution to journalArticle

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