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

TAT-WNK102 transmembrane protein and application thereof

A technology of TAT-WNK102 and transmembrane protein, which is applied in the fields of peptide/protein components, recombinant DNA technology, and medical preparations containing active ingredients, etc., and can solve problems such as unclear downstream signal transduction pathways

Inactive Publication Date: 2013-11-20
SHANGHAI UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But its downstream signal transduction pathway is not clear

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
  • TAT-WNK102 transmembrane protein and application thereof
  • TAT-WNK102 transmembrane protein and application thereof
  • TAT-WNK102 transmembrane protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Preparation of TAT-WNK102 fusion protein

[0018] The WNK102 protein corresponds to the amino acid sequence 308-409 of the human WNK-1 molecule, and is derived from a subclone using the full-length structure pEGFP-N1-hWNK-1 as a template. The fusion protein TAT-WNK102 is produced by correctly inserting the smallest translocation domain sequence (MRGSHHHHHHGMARGYGRKKRRQRRRPQ) in the Tat protein containing HIV1 into the N-terminus of the corresponding WNK1 cDNA. The fusion protein was expressed by the BL21 system in vitro and purified by the classical method using nickel affinity chromatography column.

Embodiment 2

[0019] Example 2: Culture of SD rat oligodendrocytes

[0020] Take the brains of SD newborn rats and separate the bilateral cerebral cortex, D. Rinse with Hanks solution, pipette to make cell suspension, filter with 200 mesh cell screen, centrifuge at 180×g for 5 min, discard the supernatant, add 10% fetal bovine serum in high glucose DMEM medium to resuspend the cells to (1.0 -2.0) *106 density planted in 0.025% polylysine pretreated 75c㎡ culture flask 5% CO2 culture for 8-11 days, after the cells are fused and form obvious stratification, start OPCs shaking separation and purification (the culture flask Place in a constant temperature air bath vibrating shaker, 37 degrees, 150r / min, shake for 2h to remove microglia, etc. D. Hanks solution rinse, change the medium; 5% CO2 incubator equilibrate 2h; 230r / min, 16-18h , 280r / min, 1-2h; Collect the cell suspension, centrifuge 400×g for 5 min, resuspend the cells in high glucose DMEM medium containing 10% fetal bovine serum, and plan...

Embodiment 3

[0021] Example 3: Take the SD rat oligodendrocytes purified in Example 2 and inoculate them on a 35mM culture plate pre-coated with GST-Nogo66 protein or GST protein, and replace them with fresh Neurobasal containing 2% B27 after three hours. -A culture solution (Gibco),. After culturing for 5 days, the cells were immunohistochemically performed with anti-MBP antibody and observed and photographed under a confocal microscope. see figure 1 with figure 2 The use of Nogo66 protein as a substrate to culture primary cultured rat OPC cells will indeed inhibit the differentiation and maturation of OPC cells in their differentiation medium. The number of OPC differentiated cells grown on Nogo66 substrate was significantly reduced. figure 2 for figure 1 Statistical analysis of the proportion of moderately differentiated cells. The data come from three independent experiments and are expressed as mean ± standard error. Student's-t-test was performed for comparison, p<0.001.

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 a TAT-WNK102 transmembrane protein and application thereof. The protein has an amino acid sequence shown in SEQ ID NO1. The TAT-WNK102 transmembrane protein can inhibit the combination of LINGO-1 and WNK1; and after the interaction between the LINGO-1 and the WNK1 is inhibited by the peptide, the differentiation and maturation of cultured oligodendrocytes can be promoted, thereby obtaining the TAT-WNK102 transmembrane protein. The TAT-WNK102 transmembrane protein can be used for the preparation of medicaments for treating demyelinating diseases of the central nervous system.

Description

technical field [0001] The invention relates to a TAT-WNK102 transmembrane protein and its application. Background technique [0002] In the pathogenesis of central nervous system demyelinating diseases such as multiple sclerosis, it is accompanied by the differentiation, maturation and remyelination of OPC cells. At present, the research on the molecular mechanism of how to promote the process of remyelination in patients with multiple sclerosis is a hot spot both at home and abroad. During the pathogenesis of multiple sclerosis, many inhibitory factors are produced in the disintegrated axon myelin sheath. The receptor for these inhibitors, LINGO-1, is expressed on nascent oligodendrocytes and inhibits their maturation and remyelination. The discovery of an antibody that antagonizes the function of Nogo receptor-interacting protein 1 (LINGO-1) with LRR structure and immunoglobulin domain can promote partial recovery of function in experimentally demyelinated rats. But it...

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 Patents(China)
IPC IPC(8): C12N9/12C12N15/70A61K38/45A61P25/28C12R1/19
Inventor 徐晓辉张照环黄海
Owner SHANGHAI UNIV
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