Recombined human estrogen Alpha receptor ligand binding domain protein as well as preparation and application thereof

Abstract

The invention discloses a recombined human estrogen Alpha receptor ligand binding domain protein as well as a preparation method and application of the recombined protein. The recombined protein includes a segment from the 270th amino acid residue to any one of the 554th to 595th amino acid residue of the human estrogen Alpha receptor protein as well as a 6*His tag connected with an N end of the segment and a Strep tag II connected with a C end, and is superior than the already-reported congeneric proteins in the aspects of stability and binding activity with an estrogen compound. The recombined human estrogen Alpha receptor ligand binding domain protein is provided with the double tags, thereby facilitating purification and follow-up application. An affinity adsorbent for an estrogen receptor, which is prepared by immobilizing the recombined protein on a carrier surface, can be applicable for the selective purification and the enrichment of an environment estrogen pollutant and providing a powerful way for the high throughput detection of the environment pollutant.

Description

technical field [0001] The invention relates to how to quickly and massively obtain recombinant human estrogen alpha receptor ligand binding domain protein and immobilize it on the surface of a carrier for selective purification of environmental estrogen pollutants, belonging to the field of bioanalytical chemistry. Background technique [0002] Environmental estrogen is a class of exogenous compounds that can imitate the function of biological endogenous estrogen to interfere and destroy the biological endocrine system. They are widely present in the environment and pose a serious threat to human health. In the past ten years, various analytical and detection methods have been established for various environmental estrogen substances that have been discovered, such as bisphenol A, diethylstilbestrol, alkylphenols, phthalates and organochlorine pesticides. However, these methods mainly rely on extraction and purification based on the polar characteristics of the compounds, a...

AI Technical Summary

Problems solved by technology

[0006] Judging from the composition of the recombinant human estrogen α receptor ligand-binding domain protein (rhERα-LBD) reported above, they all start from the starting amino acid 302 corresponding to the LBD or extend only 5 amino acids to the N-terminus (i.e. Starting from amino acid 297), this is not enough to protect the active conformation of LBD; in addition, from the perspective of preparation methods, the above methods only fuse a tag at one end of the protein, and multiple steps are required in the purification process, and a large amount of Protecting the activity and conformation of the receptor protein with a specific estradiol ligand is not only time-consuming and laborious, but also increases the cost; especially important, so far, no research team at home and abroad has reported the use of rhERαLBD to prepare estrogen receptors Affinity Adsorbent for Selective Purification and Enrichment of Environmental Estrogenic Pollutants

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