Molecular cloning and characterization of a secretory neutral ceramidase of Drosophila melanogaster

Yukihiro Yoshimura, Nozomu Okino, Motohiro Tani, Makoto Ito

研究成果: ジャーナルへの寄稿学術誌査読

49 被引用数 (Scopus)


We report here the molecular cloning and characterization of the Drosophila neutral ceramidase (CDase). Using the BLAST program, a neutral CDase homologue (AE003774) was found in the Drosophila GenBank and cloned from a cDNA library of Drosophila imaginal discs. The open reading frame of 2,112 nucleotides encoded a polypeptide of 704 amino acids having five putative N-glycosylation sites and a putative signal sequence composed of 23 residues. When a His-tagged CDase was overexpressed in D. melanogaster Schneider's line 2 (S2) cells, the enzyme was continuously secreted into the medium through a vesicular transport system. Treatment of the secretory 86.3-kDa CDase with glycopeptidase F resulted in the generation of a 79.3-kDa protein, indicating that the enzyme is actually glycosylated with N-glycans. The enzyme hydrolyzed various N-acylsphingosines but not galactosylceramide, GM1a or sphingomyelin, and exhibited a peak of activity at pH 6.5-7.5, and thus was classified as a neutral CDase. RNAi for the enzyme remarkably decreased the CDase activity in a cell lysate as well as a culture supernatant of S2 cells mostly at neutral pH, indicating that both activities were derived from the same gene product.

ジャーナルJournal of biochemistry
出版ステータス出版済み - 8月 2002

!!!All Science Journal Classification (ASJC) codes

  • 生化学
  • 分子生物学


「Molecular cloning and characterization of a secretory neutral ceramidase of Drosophila melanogaster」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。