秋蝇脂动激素受体的部分纯化。

N M Minnifield, D K Hayes
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引用次数: 8

摘要

神经激素受体,双翅目心体因子1和2 (DCC1和DCC2)部分纯化完成。从秋蝇(Musca autumn nalis De Geer)腹部获得受体蛋白。纯化方法包括洗涤剂增溶、亲和层析和聚丙烯酰胺凝胶电泳。凝胶电泳分析鉴定出两个相对分子量为45和90 kD的部分纯化蛋白。采用粗受体制备方法,建立了放射性标记(氚化和碘化)DCC1的配体结合试验。当过量未标记的DCC1加入到检测混合物中时,配体结合被抑制了90%。pH为7.5时配体结合最佳。结合饱和发生在约12皮摩尔放射性标记配体浓度。由于DCC1和DCC2已被证明影响昆虫的脂质和海藻糖水平,因此了解神经肽-受体相互作用对于开发新的控制乳和家禽蝇类的方法具有重要意义。
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Partial purification of a receptor for the adipokinetic hormones from Musca autumnalis face flies.

Partial purification of the receptors for the neurohormones, diptera corpora cardiaca factors 1 and 2 (DCC1 and DCC2) was achieved. Receptor proteins were obtained from the abdomens of face fly, Musca autumnalis De Geer. Purification methods included detergent solubilization, affinity chromatography, and polyacrylamide gel electrophoresis. Analysis by gel electrophoresis has identified two proteins from this partial purification with relative molecular weights of 45 and 90 kD. A crude receptor preparation was used to develop a ligand binding assay with radiolabeled (tritiated and iodinated) DCC1. Ligand binding was inhibited by 90% when excess unlabeled DCC1 was added to the assay mixture. Ligand binding was optimum at pH 7.5. Binding saturation occurred at approximately 12 picomole radiolabeled ligand concentration. Because DCC1 and DCC2 have been shown to effect the lipid and trehalose levels in the insect an understanding of the neuropeptide-receptor interaction is important for the development of new methods of control of dairy and poultry muscoid flies.

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