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Cyanea capillata polypeptide growth factor, preparation method and applications thereof

A technology of peptide growth factor and jellyfish jellyfish, which is applied in the field of jellyfish jellyfish polypeptide growth factor and its preparation, can solve the problems of easy breakage, falling off of tentacles, consumption of nematocysts, and huge number of nematocysts, etc., to achieve Significant and highly conserved effect on promoting cell proliferation

Active Publication Date: 2014-05-28
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of jellyfish preying or defending against natural enemies, on the one hand, the long and thin filamentary tentacles are prone to breakage, falling off or being bitten off by natural enemies, but the damaged parts can quickly grow new tentacles to maintain the normal state of jellyfish individuals. Activity and function, jellyfish tentacles show super damage repair ability; on the other hand, the number of nematocysts on jellyfish tentacles is huge, which needs to be produced by rapid and massive cell proliferation, and the process of predation and defense will consume a large number of nematocysts. Depleted nematocysts also need to be replenished in time, so it is speculated that there is a dynamic process of rapid proliferation of nematocysts and nematocyst replenishment in tentacles

Method used

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  • Cyanea capillata polypeptide growth factor, preparation method and applications thereof
  • Cyanea capillata polypeptide growth factor, preparation method and applications thereof
  • Cyanea capillata polypeptide growth factor, preparation method and applications thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Embodiment 1: Preparation of jellyfish nematocystoxin

[0043] Take 200g of frozen jellyfish tentacles in a beaker to thaw, add 5 times the volume of pre-cooled distilled water, and stir continuously for 10 hours with a stirrer; the suspension is filtered with a 40-mesh sampling sieve with a pore size of 450 μm, and the filtrate is collected. Centrifuge at g for 5 min, remove the supernatant, and wash the precipitate with sterile artificial seawater 3 times to obtain nematocysts; add a small amount of 50 mmol / L, pH 3.0 acetic acid to the washed nematocysts, and transfer to a medium containing the nematocysts for crushing. Put the steel balls in the crushing tube, and then use the tissue grinder Mini-Beadbeater to crush at a speed of 4600rpm for 6 minutes. Every 30s of crushing, take out the crushing tube and place it in ice water to cool for 1 minute. After the crushing is completed, centrifuge at 10000×g for 10 minutes to collect the supernatant It is jellyfish nematoc...

Embodiment 2

[0044] Embodiment 2: Separation and purification of nematocystoxin from jellyfish

[0045] (1) Superdex30 column gel filtration chromatography

[0046] The jellyfish nematocystoxin was filtered with a microporous membrane with a pore size of 0.45 μm, and the filtrate was separated and purified on a Superdex30 column, and eluted with 50 mmol / L, pH 3.0 acetic acid at a flow rate of 1 mL / min, collected and eluted by a fixed volume Peak, MALDI-TOF mass spectrometry analysis shows that the components within the outflow volume of 110-115mL (see figure 1 Elution peak SE2) contains the polypeptide growth factor Cc-GRN-1.

[0047] (2) C8 column reversed phase high performance liquid chromatography (RP-HPLC)

[0048] The above effluent volume of 110-115mL components was eluted with a linear gradient with an elution system consisting of water containing 0.1% trifluoroacetic acid and acetonitrile containing 0.1% trifluoroacetic acid, and the HPLC reversed-phase C8 column chromatography ...

Embodiment 3

[0051] Example 3: Sequencing of Cc-GRN-1 Polypeptide Growth Factor of A.

[0052] The purity of Cc-GRN-1 was identified by high performance liquid chromatography (HPLC), and its molecular weight was determined by MALDI-TOF mass spectrometry (see Figure 4 ), and its amino acid sequence was determined with an automatic protein peptide sequencer.

[0053] The jellyfish jellyfish polypeptide growth factor Cc-GRN-1 prepared by the above method is a single-chain polypeptide isolated from nematocystoxin of jellyfish jellyfish jellyfish, with a molecular weight of 5782.9Da and the primary structure of the complete amino acid sequence of the polypeptide For: Asparagine - Valine - Isoleucine - Cysteine ​​- Proline - Aspartic Acid - Glycine - Threonine - Serine - Phenylalanine - Cysteine ​​- Alanine -serine-glycine-glutamine-threonine-cysteine-cysteine-lysine-leucine-serine-serine-glycine-serine-tyrosine-glycine-cysteine- cysteine-proline-leucine-proline-asparagine-alanine-valine-cyst...

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Abstract

The invention belongs to the technical field of marine biology, and provides a cyanea capillata polypeptide growth factor named as Cc-GRN-1. The amino sequence of the growth factor is represented by the SEQ ID No. 1. The invention also provides a preparation method and applications of the cyanea capillata polypeptide growth factor. The cyanea capillata polypeptide growth factor has a prominent promoting effect on cell proliferation, and has a good application prospect in development of damage repairing drugs.

Description

technical field [0001] The invention belongs to the technical field of marine organisms, and in particular relates to a jellyfish jellyfish polypeptide growth factor and its preparation method and application. Background technique [0002] Jellyfish (jellyfish) is a kind of colloidal zooplankton, with various types, huge numbers and wide distribution, and occupies an important position in the marine ecosystem. The basic structure of jellyfish is composed of umbrella body, limbal membrane, mouth arm, tentacles, appendages, etc. The umbrella body is flat disc or spherical. There are many tentacles growing on it, the elder reaches tens of meters, and it is densely covered with nematocysts. Nematocyst is a specialized organelle of cnidocytes, a specialized cell on the tentacles. It is shaped like a small sac and contains venom. It is a defensive and offensive weapon of cnidaria. When the tentacles touch other animals, they immediately entangle the victim, and the nematocysts e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/475C07K1/20C07K1/16A61K38/18A61P17/02
CPCA61K38/00C07K14/475
Inventor 张黎明常银龙肖良郑杰民王蓓蕾王倩倩尹慢慢周永红刘丹
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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