蓝蟹血细胞海藻糖6-磷酸合酶基因的克隆、表达、酶活性及其与血淋巴海藻糖水平的关系

J Sook Chung
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引用次数: 75

摘要

海藻糖在极端环境条件下具有能量代谢和保护功能。我们从蓝蟹(Callinectes sapidus)血细胞中对海藻糖6-磷酸合成酶(TPS)进行了结构表征。与昆虫的TPS一样,C. sapidus HemoTPS (CasHemoTPS)编码TPS和海藻糖磷酸酶结构域。海藻糖似乎是一种主要的糖,因为它在血细胞和血淋巴中的含量高于葡萄糖。脂多糖刺激24 h后,血细胞中血色素TPS表达、血色素TPS酶活性和血淋巴海藻糖水平均升高,表明CasHemoTPS的TPS和TPP结构域均具有活性和功能性。TPS基因在松柏中具有广泛的组织分布,提示存在多个生物合成位点。在蜕皮周期中,血细胞中TPS活性与血淋巴海藻糖水平存在相关性。本研究首次提供了海藻糖存在于血细胞和组织中TPS的证据,并暗示其在能量代谢和生理适应中的功能作用。
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A trehalose 6-phosphate synthase gene of the hemocytes of the blue crab, Callinectes sapidus: cloning, the expression, its enzyme activity and relationship to hemolymph trehalose levels.

Trehalose in ectoderms functions in energy metabolism and protection in extreme environmental conditions. We structurally characterized trehalose 6-phosphate synthase (TPS) from hemocytes of the blue crab, Callinectes sapidus. C. sapidus Hemo TPS (CasHemoTPS), like insect TPS, encodes both TPS and trehalose phosphate phosphatase domains. Trehalose seems to be a major sugar, as it shows higher levels than does glucose in hemocytes and hemolymph. Increases in HemoTPS expression, TPS enzyme activity in hemocytes, and hemolymph trehalose levels were determined 24 h after lipopolysaccharide challenge, suggesting that both TPS and TPP domains of CasHemoTPS are active and functional. The TPS gene has a wide tissue distribution in C. sapidus, suggesting multiple biosynthetic sites. A correlation between TPS activity in hemocytes and hemolymph trehalose levels was found during the molt cycle. The current study provides the first evidence of presence of trehalose in hemocytes and TPS in tissues of C. sapidus and implicates its functional role in energy metabolism and physiological adaptation.

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