title: Molecular cloning and characterization of neutral ceramidase homologue from the red flour beetle, Tribolium castaneum.
abstract: Ceramidase plays an important role in regulating the metabolism of sphingolipids, such as ceramide, sphingosine (SPH), and sphingosine-1-phosphate (S1P), by controlling the hydrolysis of ceramide. Here we report the cloning and biochemical characterization of a neutral ceramidase from the red flour beetle Tribolium castaneum which is an important storage pest. The Tribolium castaneum neutral ceramidase (Tncer) is a protein of 696 amino acids. It shares a high degree of similarity in protein sequence to neutral ceramidases from various species. Tncer mRNA levels are higher in the adult stage than in pre-adult stages, and they are higher in the reproductive organs than in head, thorax, and midgut. The mature ovary has higher mRNA levels than the immature ovary. Tncer is localized to the plasma membrane. It uses various ceramides (D-erythro-C(6), C(12), C(16), C(18:1), and C(24:1)-ceramide) as substrates and has an abroad pH optimum for its in vitro activity. Tncer has an optimal temperature of 37 °C for its in vitro activity. Its activity is inhibited by Fe(2+). These results suggest that Tncer has distinct biochemical properties from neutral ceramidases from other species.
authors: State Key Laboratory of Rice Biology, and Molecular Biology of Crop Pathogens and Insects, The Ministry of Agriculture of China.
authors: Ying
authors: Y
authors: Zhou
authors: Xian-Wen
authors: XW
authors: Lin
authors: Qiong
authors: Q
authors: Yang
authors: Yan-Ru
authors: YR
authors: Zhang
authors: Jing-Qun
authors: JQ
authors: Yuan
authors: Xin-Da
authors: XD
authors: Lin
authors: Ruijuan
authors: R
authors: Xu
authors: Jiaan
authors: J
authors: Cheng
authors: Cungui
authors: C
authors: Mao
authors: Zeng-Rong
authors: ZR
authors: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D002518:Ceramides / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D006863:Hydrogen-Ion Concentration; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D055576:Neutral Ceramidase / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014231:Tribolium / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development
chemical_list: D002518:Ceramides; D018076:DNA, Complementary; D019476:Insect Proteins; D055576:Neutral Ceramidase