CN109517037B - Polypeptide with white spot syndrome virus inhibition effect and application thereof - Google Patents
Polypeptide with white spot syndrome virus inhibition effect and application thereof Download PDFInfo
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 36
- 241000696962 White spot syndrome virus Species 0.000 title claims abstract description 35
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 35
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- 230000000694 effects Effects 0.000 title abstract description 12
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- 230000002401 inhibitory effect Effects 0.000 claims abstract description 10
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- 230000009545 invasion Effects 0.000 abstract description 11
- 229940079593 drug Drugs 0.000 abstract description 8
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- LXAARTARZJJCMB-CIQUZCHMSA-N Ala-Ile-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LXAARTARZJJCMB-CIQUZCHMSA-N 0.000 abstract description 4
- LYILPUNCKACNGF-NAKRPEOUSA-N Ala-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C)N LYILPUNCKACNGF-NAKRPEOUSA-N 0.000 abstract description 4
- DHONNEYAZPNGSG-UBHSHLNASA-N Ala-Val-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 DHONNEYAZPNGSG-UBHSHLNASA-N 0.000 abstract description 4
- DLEBSGAVWRPTIX-PEDHHIEDSA-N Ile-Val-Ile Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)[C@@H](C)CC DLEBSGAVWRPTIX-PEDHHIEDSA-N 0.000 abstract description 4
- IWRZUGHCHFZYQZ-UFYCRDLUSA-N Phe-Arg-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 IWRZUGHCHFZYQZ-UFYCRDLUSA-N 0.000 abstract description 4
- WTWGOQRNRFHFQD-JBDRJPRFSA-N Ser-Ala-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WTWGOQRNRFHFQD-JBDRJPRFSA-N 0.000 abstract description 4
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 abstract description 3
- 231100000957 no side effect Toxicity 0.000 abstract description 3
- 150000008574 D-amino acids Chemical group 0.000 abstract description 2
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- 238000012217 deletion Methods 0.000 abstract description 2
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- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 abstract description 2
- 101710091045 Envelope protein Proteins 0.000 description 21
- 101710188315 Protein X Proteins 0.000 description 21
- 102100021696 Syncytin-1 Human genes 0.000 description 21
- 238000011160 research Methods 0.000 description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 102000005962 receptors Human genes 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 4
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- 241000238553 Litopenaeus vannamei Species 0.000 description 3
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- PEASPLKKXBYDKL-FXEVSJAOSA-N enfuvirtide Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(C)=O)[C@@H](C)O)[C@@H](C)CC)C1=CN=CN1 PEASPLKKXBYDKL-FXEVSJAOSA-N 0.000 description 1
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- 230000002194 synthesizing effect Effects 0.000 description 1
- 241000712461 unidentified influenza virus Species 0.000 description 1
- 230000007502 viral entry Effects 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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Abstract
On one hand, the invention provides a polypeptide with white spot syndrome virus inhibition effect, the amino acid sequence of which is shown as SEQ ID NO. 1 and is Ser Ala Ile Leu Ala Ile Thr Ala Val Ile Ala Val Phe Ile Val Ile Phe Arg Tyr; the amino acid sequence of the amino acid sequence is the functional equivalent of SEQ ID NO. 1, and the functional equivalent is the amino terminal or carboxyl terminal modified sequence, the sequence of D-amino acid substituted L-amino acid, and the sequence of amino acid deletion or addition of the amino acid of the SEQ ID NO. 1. The invention also provides application of the polypeptide in preparing a medicine for inhibiting white spot syndrome virus. The polypeptide obtained by the invention has the advantages of small molecular weight, easy synthesis, simple structure, no residue, no side effect, inhibition effect on the invasion of the white spot syndrome virus into prawns, and is particularly suitable for protecting prawns in the early stage of virus outbreak, and has important value in the aspect of development of environment-friendly white spot syndrome virus prevention and treatment medicines.
Description
Technical Field
The invention relates to a polypeptide with white spot syndrome virus inhibition effect, belonging to the field of biochemistry and biotechnology.
Background
White Spot Syndrome Virus (WSSV) is a main pathogenic microorganism causing large-scale death of prawns, and seriously threatens the development of the prawn breeding industry. In order to prevent and control WSSV infection, researchers make hard efforts, so far, an effective prevention and control method is still lacked, at present, the prevention and control are mainly carried out by means of breeding disease-resistant prawn varieties, strengthening epidemic disease monitoring, controlling stocking density, regularly disinfecting and the like, and the prevention and control of white spot syndrome are still far in the way.
In recent years, research shows that the specific immunity phenomenon similar to that of vertebrates exists in the prawns, and people begin immunoprophylaxis research on white spot syndrome. The vaccine prepared by recombinant Vp28 and other envelope proteins can show different degrees of protective action when being taken orally or injected for immunization. The technology is a WSSV prevention and control technology with great prospect, is still in the laboratory research stage at present, is really applied to further improving the effect of the vaccine, and depends on the deep research of prawn immune mechanisms.
Another important direction for the research of WSSV prevention and treatment is the research of virus pathogenesis, wherein the invasion mechanism is the hot point of research. Invasion of the host cell is the first step in viral infection, which is inhibited or terminated if the process can be prevented.
Binding of the virus to host cell surface receptors is a fundamental prerequisite for its invasion. Invasion of enveloped viruses relies primarily on fusion of the envelope with the host cell membrane. The envelope proteins play a key role in this process, facilitating fusion by recognizing and binding to the corresponding receptors on the cell membrane. Many viruses rely on one or two proteins to enter the host cell, and as a large enveloped virus, WSSV has more than 30 envelope proteins, of which VP28 is one of the major envelope proteins. Research shows that multiple WSSV envelope protein receptors exist on prawn cell membranes, and complex interaction exists between envelope proteins and the receptors, which is shown in that one receptor can interact with multiple envelope proteins, one envelope protein can interact with multiple receptors, and the interaction can be performed between the envelope proteins, between the envelope proteins and non-envelope proteins and between the receptors.
There is increasing evidence that the WSSV envelope proteins can form various protein complexes through different combinations, the protein complexes being the main forms of the proteins performing functions, and thus it is presumed that the WSSV envelope proteins mediate viral invasion by forming complexes.
In view of the key role of the envelope protein in the virus invasion process, the envelope protein also becomes a common target for the prevention and control research of viral diseases, and the invasion of the virus is prevented by inhibiting the action of the envelope protein. Studies have shown that polypeptides derived from the transmembrane region of the envelope protein tend to be natural inhibitors of viral entry. For example, the HIV-1 inhibiting drug Enfuvirtide is developed based on the polypeptide from HIV-1 envelope protein gb 41. In addition, influenza virus, influenza A virus, dengue virus and rift valley fever virus are screened from envelope proteins to obtain polypeptides with invasion inhibiting effect. The polypeptides have no cytotoxicity, show sequence specificity in action mode and can be used as a primer for designing virus inhibition drugs.
Because of the lack of effective WSSV control drugs at present, chemical disinfection often has the problems of residue and the like, the polypeptide provided by the invention can provide a new idea for the development of WSSV control drugs.
Disclosure of Invention
The invention aims to solve the problems of lack of ESSV control medicines and residual chemical disinfection, and provides a polypeptide with white spot syndrome virus inhibition effect, which is used for the development of WSSV inhibition medicines and has the advantages of no residue and no side effect.
The invention is realized by the following technical scheme:
on one hand, the invention provides a polypeptide with white spot syndrome virus inhibition effect, the amino acid sequence of which is shown as SEQ ID NO. 1 and is Ser Ala Ile Leu Ala Ile Thr Ala Val Ile Ala Val Phe Ile Val Ile Phe Arg Tyr; the amino acid sequence of the amino acid sequence is the functional equivalent of SEQ ID NO. 1, and the functional equivalent is the amino terminal or carboxyl terminal modified sequence, the sequence of D-amino acid substituted L-amino acid, and the sequence of amino acid deletion or addition of the amino acid of the SEQ ID NO. 1.
The invention also provides application of the polypeptide in preparing a medicine for inhibiting white spot syndrome virus.
Furthermore, the medicine is a medicine acting on the prawns.
The screening method of the polypeptide with the white spot syndrome virus inhibiting effect comprises the following steps:
(1) according to the function of envelope protein in virus invasion, selecting main envelope protein VP28 as a target from more than 20 envelope proteins of WSSV to screen polypeptide;
(2) submitting the amino acid sequence of VP28 to an MEPx server, and screening a region with a high WWIHS value in VP28 by using a hydropathic function module, namely a region with high hydrophobicity, wherein the polypeptide in the region is a candidate polypeptide;
(3) chemically synthesizing candidate polypeptide, purifying by HPLC after synthesis, and identifying by mass spectrometry;
(4) and dissolving the polypeptide by DMSO, injecting the polypeptide into prawns, injecting WSSV virus suspension at the same time, wherein the used prawns are litopenaeus vannamei, injecting only the WSSV virus suspension into a control group, and comparing the survival conditions of the two groups of prawns so as to detect the protective action of the polypeptide on the toxic-counteracting prawns.
(5) The finally screened polypeptide sequence with the WSSV inhibiting effect is as follows: ser Ala Ile Leu Ala Ile Thr Ala Val Ile Ala Val Phe Ile Val Ile Phe Arg Tyr are provided.
The polypeptide obtained by the invention has the advantages of small molecular weight, easy synthesis, simple structure, no residue, no side effect, inhibition effect on the invasion of the white spot syndrome virus into prawns, and is particularly suitable for protecting prawns in the early stage of virus outbreak, and has important value in the aspect of development of environment-friendly white spot syndrome virus prevention and treatment medicines.
Drawings
FIG. 1 is a mass spectrum of the synthetic polypeptide of example 1
FIG. 2 is a graph of the survival number and survival time of two groups of litopenaeus vannamei.
Detailed Description
The present invention will be described in further detail with reference to the following embodiments and the accompanying drawings.
Example 1
(1) Polypeptide synthesis: obtaining a polypeptide with potential inhibition effect by utilizing the hydropathiy analysis provided by the MPEx server: ser Ala Ile Leu Ala Ile Thr Ala Val Ile Ala Val Phe Ile Val Ile Phe Arg Tyr, respectively; the peptide Biotech Co., Ltd, Nanjing Jie, was assigned to the chemical synthesis by the solid phase synthesis method. Both ends of amino group and carboxyl group are not modified. After synthesis, the product was purified by HPLC to achieve a purity of 95.86% and identified by mass spectrometry, and the results are shown in FIG. 1.
(2) Control experiment: respectively arranging a toxicity attacking control group and a toxicity attacking experimental group, wherein each group contains 10 adult shrimps, and the variety of the shrimps is litopenaeus vannamei; dissolving the synthesized polypeptide in dimethyl sulfoxide (DMSO) at a concentration of 0.5 mg/mL; the virus challenge control group is injected with 40 microliters of WSSV virus suspension for each shrimp, and the virus challenge experimental group is injected with 40 microliters of polypeptide while injecting the same virus amount; and counting the survival number of the prawns every day. The results are shown in FIG. 2.
The polypeptide has obvious protection effect, particularly has obvious protection effect within the first 24 hours of virus infection (namely virus injection), the prawns in a control group die in all 4 days, and the prawns in an experimental group still survive, which indicates that the polypeptide can effectively prevent the virus from invading cells. Once the virus invades the cell, the polypeptide cannot exert a protective effect.
The above-mentioned embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solution of the present invention by those skilled in the art should fall within the protection scope defined by the claims of the present invention without departing from the spirit of the present invention.
Sequence listing
<110> Qingdao university of science and technology
<120> polypeptide having white spot syndrome virus inhibitory action and use thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 19
<212> PRT
<213> white spot syndrome virus (white spot syndrome virus)
<400> 1
Ser Ala Ile Leu Ala Ile Thr Ala Val Ile Ala Val Phe Ile Val Ile
1 5 10 15
Phe Arg Tyr
Claims (3)
1. A polypeptide having an inhibitory effect on white spot syndrome virus, characterized in that: the amino acid sequence is shown in SEQ ID NO. 1.
2. Use of the polypeptide according to claim 1 for the preparation of a medicament for inhibiting white spot syndrome virus.
3. Use according to claim 2, characterized in that: the medicine is a medicine acting on the prawns.
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| CN201811282760.0A CN109517037B (en) | 2018-10-31 | 2018-10-31 | Polypeptide with white spot syndrome virus inhibition effect and application thereof |
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| CN201811282760.0A CN109517037B (en) | 2018-10-31 | 2018-10-31 | Polypeptide with white spot syndrome virus inhibition effect and application thereof |
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| CN109517037B true CN109517037B (en) | 2022-02-08 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101166752A (en) * | 2003-09-09 | 2008-04-23 | 艾克博蒂技术公司 | Compositions and methods for inhibiting white spot syndrome virus (WSSV) infection |
| WO2009153337A1 (en) * | 2008-06-20 | 2009-12-23 | Wageningen Universiteit | Infection inhibitor |
| US7749506B2 (en) * | 1999-08-03 | 2010-07-06 | Intervet International B.V. | Proteins derived from white spot syndrome virus and uses thereof |
| CN104356210A (en) * | 2014-10-24 | 2015-02-18 | 中国水产科学研究院黄海水产研究所 | High-efficiency expression and application of VP28 protein fragment having binding activity with litopenaeus vannamei protein molecule Rab7 |
-
2018
- 2018-10-31 CN CN201811282760.0A patent/CN109517037B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7749506B2 (en) * | 1999-08-03 | 2010-07-06 | Intervet International B.V. | Proteins derived from white spot syndrome virus and uses thereof |
| CN101166752A (en) * | 2003-09-09 | 2008-04-23 | 艾克博蒂技术公司 | Compositions and methods for inhibiting white spot syndrome virus (WSSV) infection |
| WO2009153337A1 (en) * | 2008-06-20 | 2009-12-23 | Wageningen Universiteit | Infection inhibitor |
| CN104356210A (en) * | 2014-10-24 | 2015-02-18 | 中国水产科学研究院黄海水产研究所 | High-efficiency expression and application of VP28 protein fragment having binding activity with litopenaeus vannamei protein molecule Rab7 |
Non-Patent Citations (1)
| Title |
|---|
| Computational identification of self-inhibitory peptides from white spot syndrome virus envelope protein VP28;Yongzhong Lu et al.;《Aquaculture Reports》;20190415;第14卷;第1~7页 * |
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Effective date of registration: 20240204 Address after: Room 1018-15, Building 23, Times Square, Suzhou Industrial Park, Suzhou Area, China (Jiangsu) Pilot Free Trade Zone, Suzhou City, Jiangsu Province, 215000 Patentee after: Suzhou 30 billion Technology Co.,Ltd. Country or region after: China Address before: No. 53, Zhengzhou Road, North District, Qingdao, Shandong Patentee before: QINGDAO University OF SCIENCE AND TECHNOLOGY Country or region before: China |