Water-soluble membrane protein, recombinant carrier, recombinant host bacteria and its modification method and application

By modifying the interface and transmembrane amino acids of the CXCR4 receptor protein in stages, a high-affinity water-soluble membrane protein SCXCR4 was constructed, which solved the problem of reduced affinity in the existing technology and achieved highly efficient targeted intervention and nerve repair effects for cerebral hemorrhage.

CN122135767BActive Publication Date: 2026-07-14CHONGQING UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
CHONGQING UNIV
Filing Date
2026-05-07
Publication Date
2026-07-14

AI Technical Summary

Technical Problem

In existing technologies, the water-soluble modification of CXCR4 leads to a decrease in affinity with ligands, making it difficult to achieve efficient targeted intervention for cerebral hemorrhage. Furthermore, traditional biomembrane biomimetic systems have the problem of significant multi-effects but insufficient specificity.

Method used

A phased QTY Code modification strategy was adopted, based on the crystal structure of the CXCR4 receptor protein, site-directed mutagenesis interface and low-impact transmembrane amino acids, to construct a high-affinity water-soluble membrane protein SCXCR4. By screening and evaluating the water solubility and ligand binding ability of mutants, a low mutation rate and high affinity were ensured.

Benefits of technology

It maintains or enhances the binding affinity of CXCR4 and CXCL12 at a low mutation rate, achieving more efficient targeted enrichment in the cerebral hemorrhage lesion area, enhancing the blocking efficacy of the CXCL12/CXCR4 axis, and promoting neuroprotection and neurorepair.

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Abstract

The application belongs to the field of protein engineering and biological medicine, and particularly relates to a water-soluble membrane protein, a recombinant vector, a recombinant host bacterium and a modification method and application thereof. The method steps are as follows: first, interface mutants are constructed based on SQTY Code, and water solubility and ligand binding capacity are evaluated; if the requirements are not met, multiple low-impact transmembrane regions are screened out, and after mutation and modification, interface mutants are introduced to construct single-transmembrane combined mutants, and water solubility and ligand binding capacity are evaluated; if the requirements are still not met, the low-impact multiple transmembrane regions are combined two by two, and interface mutants are introduced to construct multiple double-transmembrane combined mutants, and water solubility and ligand binding capacity are evaluated, and the optimal double-transmembrane combined mutant is screened out. The method realizes water-solubilization modification of CXCR4 by rational mutation in stages, realizes water-solubilization of CXCR4, maximally changes the structure and other physicochemical properties of the protein, and further maintains or even enhances the binding capacity of the protein to the ligand CXCL12.
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