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Polypeptide for inhibiting high motor activity induced by cocaine and application thereof

A technology of polypeptides and coding genes, applied in the field of biology

Inactive Publication Date: 2011-12-14
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, existing research evidence confirms that cocaine sensitization mainly tends to activate D1 receptors in the direct loop

Method used

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  • Polypeptide for inhibiting high motor activity induced by cocaine and application thereof
  • Polypeptide for inhibiting high motor activity induced by cocaine and application thereof
  • Polypeptide for inhibiting high motor activity induced by cocaine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1, PKD1 (protein kinase D1) phosphorylation D1RC terminal specific phosphorylation site identification

[0048] 1. Preparation of GST-tagged protein

[0049] Type I dopamine receptor (hereinafter referred to as D1R) has a full length of 446 amino acids and is a typical G protein-coupled receptor with a classic seven-transmembrane structure. Therefore, the D1 receptor is segmented and defined as the N-terminus, respectively. One transmembrane domain, first intracellular loop, second transmembrane domain, first extracellular loop, third transmembrane domain, second intracellular loop, fourth transmembrane domain, second extracellular loop , the fifth transmembrane domain, the third intracellular loop, the sixth transmembrane domain, the third extracellular loop, the seventh transmembrane domain, the C-terminus ( figure 1 a).

[0050] The amino acid sequence of the type I dopamine receptor is sequence 1 in the sequence list, and the nucleotide sequence is sequenc...

Embodiment 2

[0099] Example 2, the acquisition of Tat-D1R-CT-S421 and its application in inhibiting high motor activity induced by cocaine

[0100] 1. Acquisition of Tat-D1R-CT-S421

[0101] TAT (transcriptional activator protein) penetrating peptide is one of the transmembrane delivery carriers, which can introduce covalently linked polypeptides, proteins, DNA and other molecules into cells across the membrane, and even pass through the blood-brain barrier. Without damage, it has been widely used in the fields of cell biology, gene therapy and pharmacy. According to the serine 421 of the phosphorylation site of PKD1 in D1R-CT, 14 amino acid residues including the phosphorylation preference sequence of PKD1 were selected to construct a fusion peptide with membrane penetrating ability——Tat-D1R-CT-S421. The serine at position 421 in the target sequence was mutated to alanine, ie Tat-D1R-CT-S421A was used as a control.

[0102] The amino acid sequence of Tat-D1R-CT-S421 is the sequence 2 in...

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Abstract

The invention discloses a high-motion-activity polypeptide capable of inhibiting cocaine induction and application thereof. The polypeptide disclosed by the invention is a polypeptide of (a), (b), (c) or (d) shown in the specification, wherein the (a) is a polypeptide consisting of an amino acid sequence shown in a sequence 2 in a sequence table; the (b) is a polypeptide consisting of an amino acid sequence shown as 412th to 425th amino acid residues from 5' terminal of a sequence 1 in the sequence table; the (c) is a polypeptide consisting of an amino acid sequence shown as 329th to 446th amino acid residues from the 5' terminal of the sequence 1 in the sequence table; and the (d) is a polypeptide derived from (a), (b) or (c) defined polypeptide with the same function in an amino acid sequence of the (a), (b) or (c) defined polypeptide through substitution and / or deletion and / or addition of one or more amino acid residues. Experiments of the invention prove that: the polypeptide disclosed by the invention has magnificent meaning in reducing the behavioral sensitization of a human body to cocaine stimulation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a polypeptide for inhibiting high motor activity induced by cocaine and its application. Background technique [0002] Cocaine is a very common addictive psychostimulant drug. Cocaine addiction is also a disease that seriously endangers human health and social stability. Cocaine acute stimulation, can induce increased locomotor activity. This is mainly because cocaine can inhibit the dopamine transporter in the presynaptic membrane, thereby blocking the reuptake of dopamine, resulting in an increase in the concentration of dopamine at the synaptic site. The degree of cocaine-induced locomotor activity can reflect the behavioral sensitivity of the body to cocaine, and provide some inspiration for the study of the molecular mechanism of cocaine addiction and its treatment. [0003] Dopamine stimulation can activate dopamine receptors in the postsynaptic membrane. Dopamine receptors...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N1/21C12N5/10A61K38/17A61K48/00A61P25/30C12R1/91C12R1/19
Inventor 王韵苏萍康凯王宁
Owner PEKING UNIV
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