Silkworm pupa protein source tetrapeptide SGQR and application thereof

A technology of silkworm chrysalis protein and small molecule peptides, which is applied in the field of tetrapeptide Ser-Gly-Gln-Arg, can solve the problem that the mechanism of action is not clear, and achieve the effect of inhibiting the expression level of SQS gene and protein

Active Publication Date: 2019-12-13
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at present, which peptide monomer in the silkworm chrysalis protein hydrolyzate plays a role in regulating blood lipid (cholesterol) and its mechanism of action is not clear yet.

Method used

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  • Silkworm pupa protein source tetrapeptide SGQR and application thereof
  • Silkworm pupa protein source tetrapeptide SGQR and application thereof
  • Silkworm pupa protein source tetrapeptide SGQR and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] (1) Inoculate HepG2 cells into a 6-well plate and culture for 24 hours. After discarding the old medium, add culture medium with a final concentration of 0 (blank control group) and 500ng / mL SGQR (experimental group) to each well. cultured for 12 hours respectively. The above cell culture conditions were 37°C, CO 2 Content 5%, relative humidity 95%. The medium formula is: high glucose DMEM medium containing 10% FBS, 1% penicillin / streptomycin.

[0050] (2) After the end of the culture, discard the culture medium in each well in (1), wash the cells three times with ice-cold PBS, then add 1.0ml Trizol to each well to lyse the cells, then transfer to a centrifuge tube, add 200 μl chloroform; vigorously Shake for 15Sec (avoid shaking with a shaker), let stand at room temperature for 2min, then centrifuge at 12000g for 10min at 4°C; use a pipette to draw the upper aqueous phase into another clean centrifuge tube, add 600μL of isopropanol and mix upside down, 12000g 4 Cent...

Embodiment 2

[0056] (1) Inoculate HepG2 cells in a 6-well plate and culture for 24 hours. After discarding the old medium, add culture medium with a final concentration of 0 (blank control group) and 500ng / mL SGQR (experimental group) to each well. cultured for 24 hours respectively. The above cell culture conditions are all at 37°C, with a CO2 content of 5%, and a relative humidity of 95%. The medium formula is: high glucose DMEM medium containing 10% FBS, 1% penicillin / streptomycin.

[0057] (2) Remove the culture medium from the cells in each well in (1), wash 3 times with iced PBS, add 100 μL of lysate containing protease and phosphatase inhibitors and lyse on ice for 30 minutes, scrape the cells into 1.5mL EP In the tube, heat above 95°C for 10 minutes, then centrifuge at 12,000 rpm for 10 minutes at 4°C, take the supernatant, quantify the protein, and store it in a -80°C refrigerator.

[0058] (3) Load the protein lysate in (2), first electrophoresis the stacking gel at 80V for 20m...

Embodiment 3

[0064] (1) Use the Docking module in the MOE software to carry out the molecular docking of the active peptide and HMGCR. The HMGCR structure was obtained from the crystal structure in the PDB biomacromolecular structure database (PDB code: 1HW8).

[0065] (2) Use the Docking module in the MOE software to carry out the molecular docking of the active peptide and LDLR. The LDLR structure was obtained from the crystal structure in the PDB biomacromolecular structure database (PDB code: 2MG9).

[0066] (3) Use the Docking module in the MOE software to carry out the molecular docking of the active peptide and SQS. The SQS structure was obtained from the crystal structure in the PDB biomacromolecular structure database (PDB code: 3WC9).

[0067] The molecular docking technique was used to analyze the interaction mechanism of SGQR with HMGCR, LDLR and SQS. SGQR and HMGCR can combine with four hydrogen bonds, which are the three amino acids Glu A730, Asn A734 and Glu C782 in the a...

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Abstract

The invention discloses a silkworm pupa protein source tetrapeptide SGQR. The amino acid sequence of the silkworm pupa protein source tetrapeptide SGQR is Ser-Gly-Gln-Arg. The invention also disclosesapplication of the silkworm pupa protein source tetrapeptide SGQR in preparation of lipid-lowering drugs. The SGQR can significantly reduce the HMGCR gene and protein amount, and therefore it is proved that the SGQR can reduce the synthesis amount of cholesterol.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a tetrapeptide Ser-Gly-Gln-Arg capable of inhibiting the activity of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) and lowering blood lipid (cholesterol) (SGQR). Background technique [0002] At present, the number of patients with hyperlipidemia in my country has exceeded 180 million. As my country gradually enters an aging society and people's diet and living standards improve, the number of patients with hyperlipidemia will continue to increase. The disorder of lipid metabolism, especially the disorder of cholesterol metabolism is recognized as the main factor causing cardiovascular disease. [0003] Silkworm chrysalis (bombyx mori) is a new type of healthy food raw material, which is generally raised in my country and other East Asian countries. The annual silkworm cocoons produced in my country account for 70% of the world's total cocoons. Silkworm is rich in protein and e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/103C07K1/16C12P21/06A61K38/07A61P3/06
CPCA61K38/00A61P3/06C07K5/1013C12P21/06
Inventor 孙素玲王伟张玉王君虹朱作艺李雪
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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