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New purpose of protein kinase Mnk2a

A technology of protein kinase and mnk2a, applied in peptide/protein components, medical preparations containing active ingredients, nervous system diseases, etc.

Inactive Publication Date: 2012-07-04
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Today, the kinases that can phosphorylate α-synuclein serine 129 are: casein kinases (CK1 and CK2) and G protein-coupled receptor kinases family (GRK2, GRK3, GRK5 and GRK6), etc., the mechanisms involved in the pathogenesis of PD include: ubiquitin-proteasome system, mitochondrial dysfunction and oxidative stress, etc. However, the related genes and molecular mechanisms leading to the pathogenesis of PD are still unknown. very comprehensive discussion

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  • New purpose of protein kinase Mnk2a
  • New purpose of protein kinase Mnk2a
  • New purpose of protein kinase Mnk2a

Examples

Experimental program
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Effect test

Embodiment 1

[0013] Example 1: Detection of the effect of Mnk2a on the phosphorylation of α-synuclein at position 129

[0014] 1. Construction of expression vector pcDNA3.1-α-synuclein, pcDNA3.1-α-synuclein-S129A (mutation of the serine at position 129 of α-synuclein to alanine) and pcDNA3.1-Mnk2a-myc using specific primers for PCR The α-synuclein and α-synuclein-S129A genes were cloned from fruit flies expressing human α-synuclein, and the Mnk2a gene was cloned from HEK293T cells, and then constructed into pcDNA3.1 and pcDNA3.1-myc empty loads. , Send it out for sequencing, and compare the sequencing result with the target gene sequence to confirm the success of the constructed vector.

[0015] The sequence of the upstream primer used to clone the Mnk2a target gene is: 5'-cccaagcttaccatgcccgccagccagcccattg -3'

[0016] The downstream primer sequence is: 5'-ccgctcgagtcaggcgtggtctcccaccag-3'

[0017] The full-length sequence of the Mnk2a target gene is:

[0018] atgcccgccagccagcccattgacatcccggacgc...

Embodiment 2

[0024] Example 2: Detection of the involvement of ERK signaling pathway in regulating the phosphorylation of Mnk2a on α-synuclein 129

[0025] The ERK signaling pathway activator PMA and inhibitor PD98059 were used to process cells co-transfected with the expression vectors α-synuclein and Mnk2a, and then the cell proteins were extracted for western analysis. The level of phosphorylation at serine 129 of α-synuclein in cells treated with PMA increased by 1.65 times, and the level of phosphorylation at serine 129 of α-synuclein in cells treated with PD98059 decreased by 0.80 times, and the activation of PMA could After being treated with PD98059, it decreased significantly. Experimental results indicate that Mnk2a may regulate the phosphorylation of the 129th serine of α-synuclein through the ERK signaling pathway. See results Figure 4 .

[0026] Shanghai University

[0027] New uses of protein kinase Mnk2a

[0028] 1

[0029]

[0030] 1

[0031] 1250

[0032] DNA

[0033...

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Abstract

The invention relates to a new purpose of protein kinase Mnk2a. According to the invention, the Mnk2a can be identified by a western means, the phosphorylation of Alpha-synuclein129 site serine through an extracellular signal-regulated kinase (ERK) signal passage, and at the same time, the effect that in human neuroblast oncocyte (SH-5Y-5Y) nucleuses, Mnk2a can intensify the formation of Alpha-synuclein granular aggregates similar to Lewy bodies is detected by a cell immunofluorescence means. The further basis and the action mechanism can be provided for the full study of disease generation pathogenesis of parkinson's diseases (PD).

Description

technical field [0001] The invention relates to a new application of protein kinase Mnk2a. Background of the invention [0002] Parkinson's disease (PD) is a common neurodegenerative disease of the central nervous system, and its incidence rate ranks second among neurodegenerative diseases, second only to Alzheimer's disease (senile dementia). The main pathological changes in PD patients are the degeneration and necrosis of dopamine (DA) neurons in the substantia nigra of the midbrain and the formation of Lew's body (LB) in the patient's brain. However, the mechanisms leading to the selective apoptosis of dopaminergic cells and the formation of Lewy bodies are unclear. [0003] It is generally believed that the overexpression or mutation of α-synuclein will cause the accumulation of α-synuclein to produce cytotoxicity, leading to the selective apoptosis of dopaminergic neurons, and this α-synuclein aggregate is the Lewy body (LB) in the substantia nigra. ), but what mecha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/45A61P25/16
Inventor 文铁桥夏莹
Owner SHANGHAI UNIV
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