Molecular cloning and characterization of neutral ceramidase homologue from the red flour beetle, Tribolium castaneum |
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Authors: | Zhou Ying Lin Xian-Wen Yang Qiong Zhang Yan-Ru Yuan Jing-Qun Lin Xin-Da Xu Ruijuan Cheng Jiaan Mao Cungui Zhu Zeng-Rong |
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Institution: | a State Key Laboratory of Rice Biology, China b Molecular Biology of Crop Pathogens and Insects, The Ministry of Agriculture of China, China c Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China d Department of Medicine, Medical University of South Carolina, Charleston, SC 29425, USA e Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC 29425, USA f Center for Chemical Analysis and Detection at Zhejiang University, Hangzhou, Zhejiang, China g College of Life Sciences, China Jiliang University, Hangzhou, Zhejiang 310018, China |
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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-C6, C12, C16, C18:1, and C24:1-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 Fe2+. These results suggest that Tncer has distinct biochemical properties from neutral ceramidases from other species. |
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Keywords: | Ceramidase Tncer Activity Biochemical properties Plasma membrane mRNA levels |
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