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首页> 外文期刊>Protein Expression and Purification >Efficient isolation, purification, and characterization of the Helicoverpa zea VHDL receptor
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Efficient isolation, purification, and characterization of the Helicoverpa zea VHDL receptor

机译：高效分离，纯化和表征Helicoverpa zea VHDL受体

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摘要

The study of fat body receptors (e.g., VHDL receptor) in Lepidoptera has been irksome due to the fact that isolation and purification of these proteins are difficult and resulted in extremely low yields. A rapid and efficient method is presented for the purification of Helicoverpa zea VHDL receptor by the use of VHDL biotin ligand complexed to streptavidin coated magnetic beads. The technique can be easily applied to other ligands and allows for the purification of membrane proteins with higher yields compared to previously used methods involving immunopurification. Although the purified protein can be characterized by Western and non-radioactive ligand blots using enhanced chemiluminescence (ECL), a non-radioactive ligand blot method using VHDL-FITC is presented, which allows for the quick analysis of the receptor directly from the blot under standard UV light. Sufficient receptor protein has been derived for amino acid analysis, receptor-ligand and xenobiotic binding studies. (C) 2003 Elsevier Inc. All rights reserved. [References: 13]
机译：鳞翅目中的脂肪体受体（例如，VHDL受体）的研究令人讨厌，这是因为这些蛋白质的分离和纯化困难并且导致极低的产率。提出了一种快速有效的方法，通过使用与链霉亲和素包被的磁珠复合的VHDL生物素配体来纯化Helicoverpa zea VHDL受体。与先前使用的涉及免疫纯化的方法相比，该技术可轻松应用于其他配体，并允许以更高的产率纯化膜蛋白。尽管纯化的蛋白质可以使用增强的化学发光（ECL）进行Western和非放射性配体印迹表征，但仍提出了使用VHDL-FITC的非放射性配体印迹方法，该方法可直接在以下条件下从印迹中快速分析受体标准的紫外线灯。已经获得了足够的受体蛋白用于氨基酸分析，受体-配体和异种生物结合研究。 （C）2003 Elsevier Inc.保留所有权利。 [参考：13]

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