Active peptide derived from Caprin1 and application thereof

An active peptide, stress technology, applied in the field of biomedicine

Active Publication Date: 2021-07-20
ZHEJIANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether the anticancer activity of the above small molecule compounds through the inhibition of SGs protein is dependent on the inhibition of SGs remains to be further studied to reveal

Method used

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  • Active peptide derived from Caprin1 and application thereof
  • Active peptide derived from Caprin1 and application thereof
  • Active peptide derived from Caprin1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Design of polypeptide SGs inhibitors based on blocking the interaction between Caprin1 and G3BP1

[0043] 1. According to research reports, the interaction between the two nucleating proteins Caprin1 of SGs and G3BP1 will promote the assembly of SGs. The present invention explores the minimum G3BP1 binding region of the Caprin1 protein, and designs four polypeptide fragments based on the Caprin1 protein: Caprin1(258-415), Caprin1(301-390), Caprin1(301-350), and Caprin1(351-390). Entrust Shanghai Gil Biochemical Company to synthesize the above polypeptide fragments.

[0044] 2. Construct Caprin1 and hG3BP1 expression plasmids, design and synthesize the following primers:

[0045] G3BP1:

[0046] G3BP1-F 5'-atggtgatggagaagcctagtcccctgct-3' (SEQ ID No: 14);

[0047] G3BP1-Xba-R 5'-ccatctagattcactgccgtggcgcaagcc-3' (SEQ ID No: 15);

[0048] Caprin1:

[0049] Caprin1-F 5'-atgccctcggccaccagccaca-3' (SEQ ID No: 16);

[0050] Caprin1-Xba-R 5'-tcttctagaaacatattat...

Embodiment 2

[0058] Example 2 investigates the ability of the active peptide derived from Caprin1 of the present invention to compete with the protein Caprin1 for binding to G3BP1

[0059] Based on the research results of Example 1, Caprin1 (361-385) with a shorter sequence was designed on the basis of Caprin1 (351-390).

[0060] 1. Construction of Caprin1(361-385), Caprin1(351-390) expression plasmids

[0061] The following primers were designed and synthesized:

[0062] Caprin1 (351-390):

[0063] Caprin1-351F 5'-gatccccttgtgagaagacagcgagtacaa-3' (SEQ ID No: 18);

[0064] Caprin1-Xba-390R 5'-ctgtctagatacaatggcaggatcaagtgt-3' (SEQ ID No: 19);

[0065] Caprin1 (361-385):

[0066] Caprin1-361F 5'-gaccttatggcacaaatgcagggtccctat-3' (SEQ ID No: 20);

[0067] Caprin1-Xba-385R 5'-tgatctagaaagtgtctgattttcaaaatc-3' (SEQ ID No: 21).

[0068] Referring to "Molecular Cloning Experimental Guide (Fourth Edition)" (Cold Spring Harbor Laboratory Press / Science Press), use the pCS2-Myc-hCaprin1 prepa...

Embodiment 3

[0083] Example 3 To investigate the activity of Caprin1(361-385) and Caprin1(351-390) in inhibiting SGs HeLa cell line was seeded in 6-well plate, and when the cell confluency reached about 60%, the expression of Caprin1 constructed in Example 2 was carried out. (361-385) and Caprin1(351-390) plasmids were transfected into HeLa cells and overexpressed in the cells. When the cell confluency reaches about 80%, take 12 mL of cell culture base into a 15 mL centrifuge tube, add 12 μL of 0.5 mM AS, mix well, replace the medium in the 6-well plate, and stimulate HeLa cells to form SGs (use endogenous G3BP1 as the SGs Mark). Put into the carbon dioxide cell incubator and continue to cultivate for 45min. Remove the 6-well plate for immunofluorescence experiments.

[0084] The result is as image 3 As shown, the results indicated that overexpression of Caprin1(351-390), Caprin1(361-385) inhibited the SGs induced by AS compared with the control group ( image 3 A). In order to ensur...

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Abstract

The invention designs an active peptide derived from Caprin1, which can be competitively combined with the G3BP1 protein to specifically block the interaction between the Caprin1 and the G3BP1 protein and inhibit generation of stress particles so as to treat diseases related to the generation of SGs, such as tumors or neurodegenerative diseases.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a class of active peptides derived from Caprin1, which can competitively bind G3BP1 / 2 with Caprin1, inhibit the assembly of stress granules, and produce corresponding biological effects. Background technique [0002] There are many non-membrane structures rich in RNA and protein in cells, called ribonucleoprotein (RNP). These RNP granules are individual organelles composed of thousands of molecules and are ubiquitous in cells. When eukaryotic cells respond to stress stimuli such as endoplasmic reticulum stress, heat shock, hypoxia, and arsenate, they will form aggregated granules of mRNA-protein complexes in the cytoplasm, namely stress granules. SGs). [0003] The formation of SGs has important physiological significance, including the regulation of mRNA translation and the inhibition of apoptosis-related signaling pathways, to minimize the damage caused by stress to cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K19/00C12N15/12A61K38/17A61P35/00A61P9/10A61P25/00A61P25/08A61P25/28A61P25/16A61P25/14
CPCC07K14/4702A61P35/00A61P9/10A61P25/00A61P25/08A61P25/28A61P25/16A61P25/14C07K2319/10A61K38/00
Inventor 金志刚李娟周钰林金博星李晨辉
Owner ZHEJIANG NORMAL UNIVERSITY
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