Polypeptides with function of self-assembled potassium channel and its application

A potassium channel and self-assembly technology, applied in the field of polypeptides with self-assembled potassium channel function, can solve the problem of lack of synthetic ion channel drugs, etc., and achieve the effects of high purity, small molecular weight and low cost

Inactive Publication Date: 2013-03-27
ANHUI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention is mainly aimed at the lack of synthetic ion channel drugs in the field of bioengineering drug development at present, and uses natural amino acid sequences to develop synthetic ion channel drugs, and provides a polypeptide with self-assembled potassium ion channels on cell membranes and its application

Method used

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  • Polypeptides with function of self-assembled potassium channel and its application
  • Polypeptides with function of self-assembled potassium channel and its application
  • Polypeptides with function of self-assembled potassium channel and its application

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preparation example Construction

[0024]The preparation method of the polypeptide of the present invention adopts a chemical synthesis method, that is, a very mature solid-phase peptide synthesis method well known in the art, and its amino protection strategies mainly include 9-fluorenylmethoxycarbonyl (Fmoc) method and tert-butoxycarbonyl (Boc ) method two, at present domestic and foreign mainly adopt Fmoc solid-phase synthesis method. The solid-phase synthesis method can use commercially available protected amino acids as raw materials to obtain a large number of samples quickly and efficiently. In the 9-fluorenylmethoxycarbonyl (Fmoc) synthesis system, preferably Wang (Wang) resin, add 15ml / g of dimethylformamide (DMF) for 30 minutes, then add a total of 20% of hexahydropyridine to treat at room temperature 20 minutes to remove the Fmoc protecting group, and then add an excess of the corresponding protected amino acid and the activator benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), ...

Embodiment 1

[0037] A polypeptide with self-assembling potassium channel function is a polypeptide containing 24 amino acids: the primary structure of the complete sequence of the polypeptide is: leucine-tyrosine-leucine-alanine-valine-formazine Thionine-Valine-Phenylalanine-Alanine-Leucine-Valine-Leucine-Glycine-Tryptophan-Methionine-Asparagine-Alanine-Leucine Acid-tyrosine-phenylalanine-threonine-arginine-glycine-leucine (the amino acid sequence of the polypeptide is: Leu-Tyr-Leu-Ala-Val-Met-Val-Phe-Ala -Leu-Val-Leu-Gly-Trp-Met-Asn-Ala-Leu-Tyr-Phe-Thr-Arg-Gly-Leu).

[0038] Polypeptide of the present invention (Leu-Tyr-Leu-Ala-Val-Met-Val-Phe-Ala-Leu-Val-Leu-Gly-Trp-Met-Asn-Ala-Leu-Tyr-Phe-Thr-Arg-Gly- Leu) solid-phase synthesis

[0039] The polypeptide of the present invention whose sequence is LYLAVMVFALVLGWMNALYFTRGL was synthesized by a commercially available Quartet polypeptide synthesizer (PTI company), and the specific steps were as follows:

[0040] 1. Add appropriate amount...

Embodiment 2

[0047] Polypeptide of the present invention (Leu-Tyr-Leu-Ala-Val-Met-Val-Phe-Ala-Leu-Val-Leu-Gly-Trp-Met-Asn-Ala-Leu-Tyr-Phe-Thr-Arg-Gly- Leu) forms K on lipid bilayer membranes + ion single channel current

[0048] 1. Materials and methods

[0049] 1.1 Materials: KCl and NMDG-Cl were purchased from Sigma Company, POPC phospholipids and phosphatidylserine (phosphatidylserine, PS) were purchased from Avanti Company (Avanti Polar Lipids, USA), inverted microscope (IX70, Olympus, Japan), and microelectrodes were drawn instrument (P-97, Sutter, USA), three-dimensional microelectrode manipulator (MP-285, Sutter, USA), patch clamp amplifier (EPC-9, HEKA, Germany), PULSE+PULSEFIT recording and analysis software (HEKA, Germany).

[0050] 1.2 Synthesis of lipid bilayers: Weigh 80 mg POPC and 20 mg phosphatidylserine, dissolve them in 2 ml double distilled water, suspend with a vortex mixer for 20 minutes under nitrogen, and then ultrasonically emulsify for 10 minutes. Centrif...

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Abstract

The invention provides polypeptides with the function of self-assembled potassium channel. The main sequence of polypeptides fragmentations, analog, derivative and variant amino acid of polypeptide possesses great than or equal to 70% of homology and great than or equal to 90% of similarity compared with the main sequence of the polypeptides amino acid. Polypeptides, nucleotide, polypeptide fragmentations, polypeptide analog, polypeptide derivatives and polypeptides variant are used for preparing the medicines which can treat diseases caused by potassium channel functional disorder / deficiency and hypertension, and also used for preparing tool drugs in the research field which can modify liposome and cell membrane potential. The invention provides a novel method for treating diseases caused by potassium channel functional disorder / deficiency and modifying cell membrane potential.

Description

technical field [0001] The invention belongs to the field of bioengineering drug development, and specifically relates to a polypeptide with self-assembled potassium channel function and application thereof. Background technique [0002] In the process of life activities, due to self-metabolism and response to various physiological stimuli, the ion concentration in the cell often changes, and the main way to regulate the ion concentration in the cell is the ion channel on the cell membrane. Ion channels play an important role in many physiological responses, such as: the generation and transmission of electrical signals in the nervous system, muscle contraction, gland secretion, cytoplasmic pH regulation, cardiovascular system function, immune response, etc. Ion channels are generally membrane proteins composed of several identical or different subunits. Each subunit contains several hydrophobic transmembrane segments. The transmembrane segments of different subunits surroun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/00C12N15/11A61K38/16A61K48/00A61P9/12A61P43/00
Inventor 沈兵杜鹃范礼斌柯道平朱金行
Owner ANHUI MEDICAL UNIV
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