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Polypeptide for inhibiting actin recombination and application thereof

A technology of actin and preparation, which is applied in the field of peptides and applications that inhibit actin recombination, and achieves the effect of weak immunogenicity and strong specific binding ability

Active Publication Date: 2019-09-03
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there is no polypeptide drug that competitively inhibits the combination of α-actinin and actin to inhibit the recombination of F-actin at home and abroad

Method used

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  • Polypeptide for inhibiting actin recombination and application thereof
  • Polypeptide for inhibiting actin recombination and application thereof
  • Polypeptide for inhibiting actin recombination and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] HIV-1 entry into CD4 by ABS1p + Inhibitory effect of T cells or CEM-ss cells

[0029] Studying the effect of ABS1p on HIV-1 entry into CD4 + For the inhibitory effect of T cells or CEM-ss cells, first adjust the cell concentration to 1×10 6 cells / mL, the volume of each group of cells is 2mL; add peptide ABS1p to the cell system, and incubate in a 37°C incubator for 1h; after pretreatment, add 200ng p24HIV-1 to the cells, place at 37°C, 5% CO 2 After the infection was completed, the cells were rinsed with PBS for 3 times, and fresh medium was added to make the cell concentration 1×10 6 cells / mL, keep at 37°C, 5% CO 2 Conditioned for 48h.

[0030]When measuring the infection effect of the virus on the cells, Intracellular p24staining was used to detect the p24 content in the cells after virus infection. In the experiment, the virus-infected cells were first collected, washed with PBS for 3 times, and then p24 in the cells were stained according to the Anti-HIV-1p24an...

Embodiment 2

[0033] ABS1p affects the recombination of F-actin and α-actinin in T cells:

[0034] order in CD4 + To study the dynamic relationship between F-actin and α-actinin in T cells and T cell line CEM-ss cells, co-transfect CD4 with plasmid pECFP-C1-lifeact and piRFP-α-actinin + T cells or CEM-ss cells can achieve fluorescent labeling of F-actin and α-actinin in living cells. The specific implementation process is: (1) add 100 μ L room temperature Solution resuspended CD4 + Add 2 μg pECFP-C1-Lifeact and 2 μg piRFP-α-actinin to T cells or CEM-ss cells, and mix gently; (2) Add the cell plasmid mixture into the small groove of the electroporation cup, and cover the cap of the electroporation cup; ( 3) Set the program of the LONZA nuclear transfer instrument: select the V-024 program, insert the electro-transfer cuvette containing the cells and plasmids into the slot of the nuclear transfection instrument, and run the selected program; (4) After the program is finished, immediately ...

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Abstract

The invention discloses a polypeptide for inhibiting actin recombination and an application thereof. The polypeptide has the amino acid sequence shown in SEQ ID NO.1 and the nucleotide sequence of a gene encoding ABS1p is shown in SEQ ID NO.2. ABS1p is based on ABS of an ABD sequence of non-muscular alpha-actinin and can bind to F-actin in primary CD4+T cells. Experiments confirm that the ABS1p inhibits the recombination of the T cell F-Actin and the infection of HIV-1 to the T cells and is expected to develop into a functional inhibitor for AIDS treatment, or can be combined with other inhibitors to achieve an ideal therapeutic effect.

Description

technical field [0001] The invention belongs to the technical field of biomedical engineering, and specifically relates to a polypeptide for inhibiting actin recombination and its application. The polypeptide provided by the invention inhibits HIV-1 infection by inhibiting actin recombination. Background technique [0002] Human immunodeficiency virus (Human Immunodeficiency Virus, HIV) is a lentivirus that infects cells of the human immune system. It belongs to the family Retroviridae and the genus Lentivirus. AIDS), namely AIDS. It can cause damage to the immune system of the body and pose a great threat to human health. [0003] At present, anti-AIDS drugs are mainly divided into four categories: reverse transcriptase inhibitors, protease inhibitors, integrase inhibitors and entry inhibitors. Unlike the first three classes of drugs, entry inhibitors target the early stages of viral infection. According to the three steps that drugs target HIV to enter cells, entry inhi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/12A61K38/17A61P31/18
CPCA61K38/00C07K14/47
Inventor 崔宗强印文李炜张先恩李芹刘霁
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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