Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Fusion peptide TAT-MAS9C for destroying interaction between Mas receptors and PSD95

A TAT-MAS9C, fusion peptide technology, applied in the field of fusion peptides, can solve the problems of inappropriate PSD95 regulation function research, low neuronal transfection efficiency, etc., and achieve the effect of strengthening the protective effect of cerebral ischemia

Inactive Publication Date: 2016-08-03
BINZHOU MEDICAL COLLEGE
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Since both Mas and PSD95 are expressed in nerve cells, but the transfection efficiency of nerve cells is very low, and PSD95 itself has a certain synaptic function, so the method of changing the expression of PSD95 to study the regulation of PSD95 on Mas is not only helpful. Technically difficult, and not suitable for research on the regulatory function of PSD95

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Fusion peptide TAT-MAS9C for destroying interaction between Mas receptors and PSD95
  • Fusion peptide TAT-MAS9C for destroying interaction between Mas receptors and PSD95
  • Fusion peptide TAT-MAS9C for destroying interaction between Mas receptors and PSD95

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Design and synthesis of the fusion peptide of the present invention

[0032] The fusion peptide TAT-MAS9C used to destroy the interaction between the Mas receptor and PSD95 has a full length of 20 amino acids, and its sequence includes two parts. The amino terminal of the fusion peptide is the transmembrane domain of the HIV-1 Tat protein. The 11 amino acids YGRKKRRQRRR can introduce the fusion peptide into the cytoplasm; the carboxy-terminal of the fusion peptide is the 9 amino acids NTVSIETVV at the carboxy-terminal of Mas, which can destroy the interaction between Mas and PSD95. The amino acid sequence of the fusion peptide is YGRKKRRQRRRNTVSIETVV.

[0033] For some research needs, rhodamine can be attached to amino acid K of the fusion peptide for fluorescent labeling. The method for producing the fusion peptide is well known, and general peptide synthesis companies can synthesize it by solid-phase synthesis.

[0034] The method for synthesizing the fu...

Embodiment 2

[0036] Example 2 Verification of the transmembrane effect of TAT-MAS9C by fluorescence microscopy

[0037] Program:

[0038] The experiment was divided into two groups: group A was rhodamine-labeled negative control group; group B was TAT-MAS9C group. Primary cultured neurons for 7 days, after the synapse grows, use rhodamine-labeled negative control and 10 -5 M's TAT-MAS9C incubated neurons for 30 minutes, washed away the negative control and TAT-MAS9C, and took pictures with a fluorescence microscope to observe their transmembrane effects.

[0039] result:

[0040] After washing off the peptide, it was found that the negative control in group A had a poor penetration effect and the fluorescence intensity was very weak ( Figure 1A ). In the cytoplasm of neurons in group B, a large number of fluorescently labeled TAT-MAS9C ( Figure 1B ), indicating that TAT-MAS9C has a good transmembrane effect.

Embodiment 3

[0041] Example 3 The interference effect and specificity of TAT-MAS9C were verified by CoIP (co-immunoprecipitation) combined with western blot (immuno-western blotting) method:

[0042] Program:

[0043] The experiment was divided into three groups. The 293 cells were seeded in 90mm cell culture dishes, and when the density was about 70%, the 293 cells were transfected with Flag-Mas and PSD95 plasmids at the same time. 24 hours after transfection, add the negative control to the culture medium of group A and incubate for 30 minutes; add the final concentration of 10 -5 M's TAT-MAS9C, incubated for 30min; C group added 10 -6 M TAT-MAS9C, incubated for 30min. Cells were harvested with protein lysate, mixed continuously at 4°C for 1 h, and the supernatant was collected by centrifugation. Add 50 μL of Anti-FlagM2AffinityGel to 1 mL of supernatant, and keep mixing for 3 hours at 4°C. Centrifuge for 1 min to collect the precipitate, wash the precipitate with washing solution,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to fusion peptide TAT-MAS9C for destroying interaction between Mas receptors and PSD95. The full-length fusion peptide TAT-MAS9C comprises 20 amino acids; the sequence of the fusion peptide TAT-MAS9C comprises two parts; the amino terminal of the fusion peptide comprises 11 amino acids YGRKKRRQRRR in a membrane spanning domain of HIV-1Tat protein and can be used for importing the fusion peptide into cytoplasm; the carboxyl terminal of the fusion peptide comprises 9 Mas carboxy-terminal amino acids NTVSIETVV and can be used for destroying the interaction between the Mas and the PSD95; the amino acid sequence of the fusion peptide is YGRKKRRQRRRNTVSIETVV. The fusion peptide provided by the invention is capable of destroying the interaction between the Mas and the PSD95 and ensuring the synaptic function of PSD95, and can be used for preferably researching the regulating effect of the PSD95 on the Mas function.

Description

technical field [0001] The present invention relates to a fusion peptide for disrupting the interaction between Mas receptor and PSD95. Background technique [0002] The mas gene was first cloned in 1986 by Dr. Wigler's research group. The protein encoded by it contains 325 amino acids, has 7 hydrophobic transmembrane domains, and is a G protein-coupled receptor. The histological distribution of Mas protein is relatively extensive, and it is expressed in brain, testis, heart, kidney and cerebral vascular endothelial cells. Mas has a variety of physiological functions, it can effectively regulate the renin-angiotensin system; at the same time, it also has an important regulatory effect on neuroplasticity, memory and anxiety; in addition, it also has an important impact on the occurrence of tumors. The current research on Mas focuses on the function and signaling pathway, and there are few research reports on Mas regulatory proteins. [0003] The carboxy-terminal (ETVV) of ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C07K1/06C07K1/04A61K38/00A61P25/00A61P9/10
CPCY02P20/55C07K14/705A61K38/00C07K14/00C07K2319/00
Inventor 卞伟华
Owner BINZHOU MEDICAL COLLEGE
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com