A metal-affinity fusion protein tag and its application

Abstract

The invention discloses a novel metal-affinity fusion protein tag, which is derived from the extracellular N-terminal fragment of a natural human copper ion transporter, which can replace His-tag as a metal-affinity tag for purifying recombinant fusion proteins; Also disclosed is a novel fusion protein fused with the tag, which has special physiological and pharmacological activities.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a novel metal affinity fusion protein tag with affinity for metal ions, and a fusion protein thereof. Background technique [0002] Metal chelation affinity chromatography, also known as immobilized metal ion affinity chromatography. The method utilizes the metal ion binding properties of some amino acids on the protein surface, such as histidine, tryptophan and other residues, and separates and purifies the protein through the adsorption of the solid phase matrix. Histidine tag (His-Tag), as a synthetic sequence, can be used for fusion expression of exogenous protein to facilitate the purification of recombinant protein. At the amino-terminus, carboxyl-terminus or middle of the target protein, a peptide segment consisting of continuous histidine residues (usually 6-10 histidines) can be fused in a variety of expression systems (bacteria, yeast and mammalian cells) suc...

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Problems solved by technology

[0003] It has been found that the fusion of His-tag at the amino or carboxyl terminal of the protein will affect the renaturation[1], stability[2] or solubility[3] of the protein; His-tag may also affect the activity of the protein, such as His -tag is fused to the amino terminal of Chlorocatchol1,2-Dioxygenase (CCD), although its structure is only slightly different from CCD, its activity is reduced by 5 times[4]

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