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从黄热病蚊子埃及伊蚊中分离并表达一种固醇载体蛋白-2基因。

Isolation and expression of a sterol carrier protein-2 gene from the yellow fever mosquito, Aedes aegypti.

作者信息

Krebs K C, Lan Q

机构信息

Department of Entomology, University of Wisconsin-Madison, Madison, WI 53076, USA.

出版信息

Insect Mol Biol. 2003 Feb;12(1):51-60. doi: 10.1046/j.1365-2583.2003.00386.x.

Abstract

Trafficking of cholesterol in insects is a very important process due to the fact that insects depend on dietary cholesterol to fulfil their physiological needs. We identified a putative mosquito sterol carrier protein-2 (SCP-2) cDNA from fourth instar subtracted cDNA library. The AeSCP-2 protein has high degree homology in the sterol transfer domain to both rat and human SCP-2. Transcripts of AeSCP-2 in fourth instars were detected strongly in the midgut, and weakly in the head and hindgut. In the early pupae, AeSCP-2 transcription was observed in the thorax, head and body wall of abdomen, but not in the gut. The interaction of mosquito sterol carrier protein-2 (AeSCP-2) with cholesterol was examined. The Kd of purified recombinant AeSCP-2 to cholesterol was 5.6 +/- 0.6 x 10-9 m using radiolabelled cholesterol-binding assay. The results suggest that AeSCP-2 has high affinity to cholesterol and may function as a carrier protein in mosquitoes.

摘要

由于昆虫依赖膳食胆固醇来满足其生理需求,因此胆固醇在昆虫体内的运输是一个非常重要的过程。我们从四龄幼虫消减cDNA文库中鉴定出一个推定的蚊子固醇载体蛋白2(SCP-2)cDNA。埃及伊蚊SCP-2蛋白在固醇转移结构域与大鼠和人类SCP-2具有高度同源性。在四龄幼虫中,埃及伊蚊SCP-2的转录本在中肠中检测到的信号很强,而在头部和后肠中检测到的信号较弱。在早期蛹中,在胸部、头部和腹部体壁中观察到埃及伊蚊SCP-2的转录,但在肠道中未观察到。研究了蚊子固醇载体蛋白2(AeSCP-2)与胆固醇的相互作用。使用放射性标记的胆固醇结合试验,纯化的重组AeSCP-2对胆固醇的解离常数(Kd)为5.6±0.6×10-9 m。结果表明,AeSCP-2对胆固醇具有高亲和力,可能在蚊子中作为载体蛋白发挥作用。

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