A neutral ceramidase homologue from Dictyostelium discoideum exhibits an acidic pH optimum

Hatsumi Monjusho, Nozomu Okino, Motohiro Tani, Mineko Maeda, Motonobu Yoshida, Makoto Ito

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Ceramidases (CDases) are currently classified into three categories (acid, neutral and alkaline) based on their optimal pHs and primary structures. Here, we report the first exception to this rule. We cloned the CDase cDNA, consisting of 2142 nucleotides encoding 714 amino-acid residues, from the slime mould, Dictyostelium discoideum. The putative amino-acid sequence indicates 32-42% identity with various neutral CDases, but does not show any similarity to the acid and alkaline CDases, indicating the enzyme should be classified as a neutral CDase. However, overexpression of the cDNA in D. discoideum resulted in increased CDase activity at an acidic, but not a neutral pH range. Knockout of the gene in slime mould eliminated CDase activity at acidic pH. The recombinant enzyme expressed in the slime mould was purified and then characterized. Consequently, the purified CDase was found to exhibit the maximal activity at approx. pH 3.0. The singular pH dependency of slime mould CDase is not derived from the specific post-translational modification in the slime mould, because the enzyme showed an acidic pH optimum even when expressed in Chinese hamster ovary cells, whereas rat neutral-CDase exhibited a neutral pH optimum when expressed in slime mould.

Original languageEnglish
Pages (from-to)473-479
Number of pages7
JournalBiochemical Journal
Volume376
Issue number2
DOIs
Publication statusPublished - Dec 1 2003

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Neutral Ceramidase
Dictyostelium
Fungi
Alkaline Ceramidase
Acid Ceramidase
Ceramidases
Enzymes
Complementary DNA
Amino Acids
Rats
Nucleotides
Genes
Cells
Gene Knockout Techniques
Acids
Post Translational Protein Processing
Cricetulus
Amino Acid Sequence
Ovary

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A neutral ceramidase homologue from Dictyostelium discoideum exhibits an acidic pH optimum. / Monjusho, Hatsumi; Okino, Nozomu; Tani, Motohiro; Maeda, Mineko; Yoshida, Motonobu; Ito, Makoto.

In: Biochemical Journal, Vol. 376, No. 2, 01.12.2003, p. 473-479.

Research output: Contribution to journalArticle

Monjusho, Hatsumi ; Okino, Nozomu ; Tani, Motohiro ; Maeda, Mineko ; Yoshida, Motonobu ; Ito, Makoto. / A neutral ceramidase homologue from Dictyostelium discoideum exhibits an acidic pH optimum. In: Biochemical Journal. 2003 ; Vol. 376, No. 2. pp. 473-479.
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