Molecular cloning and characterization of sphingolipid ceramide N-deacylase from a marine bacterium, Shewanella alga G8

Masako Furusato, Noriyuki Sueyoshi, Susumu Mitsutake, Keishi Sakaguchi, Katsuhiro Kita, Nozomu Okino, Sachiyo Ichinose, Akira Omori, Makoto Ito

研究成果: ジャーナルへの寄稿記事

16 引用 (Scopus)

抄録

Recently, lyso-sphingolipids have been identified as ligands for several orphan G protein-coupled receptors, although the molecular mechanism for their generation has yet to be clarified. Here, we report the molecular cloning of the enzyme, which catalyzes the generation of lyso-sphingolipids from various sphingolipids (sphingo-lipid ceramide N-deacylase). The 75-kDa enzyme was purified from the marine bacterium, Shewanella alga G8, and its gene was cloned from a G8 genomic library using sequences of the purified enzyme. The cloned enzyme was composed of 992 amino acids, including a signal sequence of 35 residues, and its molecular weight was estimated to be 109,843. Significant sequence similarities were found with an unknown protein of Streptomyces fradiae Y59 and a Lumbricus terrestris lectin but not other known functional proteins. The 106-kDa recombinant enzyme expressed in Escherichia coli hydrolyzed various glycosphingolipids and sphingomyelin, although it seems to be much less active than the native 75-kDa enzyme. In vitro translation using wheat germ extract revealed the activity of a 75-kDa deletion mutant lacking a C terminus to be much stronger than that of the full-length enzyme, suggesting that C-terminal processing is necessary for full activity.

元の言語英語
ページ(範囲)17300-17307
ページ数8
ジャーナルJournal of Biological Chemistry
277
発行部数19
DOI
出版物ステータス出版済み - 5 10 2002

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Shewanella
Ceramides
Cloning
Molecular Cloning
Algae
Bacteria
Lipids
Enzymes
Glycosphingolipids
Oligochaeta
Genomic Library
Sphingomyelins
Streptomyces
Protein Sorting Signals
G-Protein-Coupled Receptors
Lectins
Escherichia coli
Triticum
Proteins
Genes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Molecular cloning and characterization of sphingolipid ceramide N-deacylase from a marine bacterium, Shewanella alga G8. / Furusato, Masako; Sueyoshi, Noriyuki; Mitsutake, Susumu; Sakaguchi, Keishi; Kita, Katsuhiro; Okino, Nozomu; Ichinose, Sachiyo; Omori, Akira; Ito, Makoto.

:: Journal of Biological Chemistry, 巻 277, 番号 19, 10.05.2002, p. 17300-17307.

研究成果: ジャーナルへの寄稿記事

Furusato, Masako ; Sueyoshi, Noriyuki ; Mitsutake, Susumu ; Sakaguchi, Keishi ; Kita, Katsuhiro ; Okino, Nozomu ; Ichinose, Sachiyo ; Omori, Akira ; Ito, Makoto. / Molecular cloning and characterization of sphingolipid ceramide N-deacylase from a marine bacterium, Shewanella alga G8. :: Journal of Biological Chemistry. 2002 ; 巻 277, 番号 19. pp. 17300-17307.
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