Protein and small molecule polypeptide for increasing lysozyme bacteriolysis activity and its use

A protein molecule, lysozyme technology, applied in the field of TRAP protein, homologous polypeptide molecules, can solve the problem of no functional activator

Inactive Publication Date: 2007-04-11
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no research report on its functional activator
There is no research report on other functions of TRAP protein

Method used

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  • Protein and small molecule polypeptide for increasing lysozyme bacteriolysis activity and its use
  • Protein and small molecule polypeptide for increasing lysozyme bacteriolysis activity and its use
  • Protein and small molecule polypeptide for increasing lysozyme bacteriolysis activity and its use

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0013] 1. Binding of TRAP protein to lysozyme and activation of lysozyme activity

[0014] 1.1 Materials

[0015] TRAP protein was expressed and purified by our laboratory, and lysozyme was purchased from SIGMA. Polyclonal antibodies to TRAP and lysozyme were prepared by our laboratory. HRP-labeled anti-rabbit secondary antibody and lysozyme activity kit were purchased from Laibo Company.

[0016] 1.2 Method

[0017] 1.2.1 ELISA method to detect the combination of TRAP protein and lysozyme

[0018] ① Coat the ELISA plate with TRAP protein at a coating concentration of (2 μg / well), overnight at 4°C, wash once with PBST, block with 10% fetal bovine serum at 37°C for 1 hour, wash with PBST three times, add lysozyme (5 μg / well) Wells), incubated at 37°C for 1 hour, washed three times with PBST, added lysozyme antibody (1:1000), incubated at 37°C for 1 hour, washed three times with PBST, added HRP-labeled anti-rabbit secondary antibody (1:10000), TMB Substrate chromogenic solu...

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Abstract

The present invention is TRAP protein capable of combining with lysozyme mutually and promoting the activity of lysozyme and its homologous small molecular polypeptides. The polypeptides possess amino acids with the general expression of S / T-Y / W-F-E / D-X-X-L-Y-P / A-X-X-X, where S represents serine residue, T threonine, Y asparatic acid residue, W tryptophane residue, F phenylalanine residue, E glutamic acid residue, D aspartic acid residue, L leucine residue, P proline residue, A alanine residue, X any natural L-amino acid residue or D-isomer. The TRAP protein and the polypeptides of the present invention may be applied in medicine and microbe fermentation fields for enhancing the bacteriolysis activity of lysozyme.

Description

technical field [0001] The invention relates to a TRAP protein and a group of homologous polypeptide molecules capable of combining with lysozyme and promoting the activity of lysozyme. Background technique [0002] Lysozyme is known to kill cells by breaking down the cell wall on the surface of bacteria. Lysozyme from various sources has been used in clinical research. But there is no research report about its functional activator. TRAP protein is a surface protein of Staphylococcus aureus, and its main biological function is to participate in the regulation of Staphylococcus aureus toxin. With the increase of TRAP protein phosphorylation level, the exotoxin level of Staphylococcus aureus is up-regulated (Balaban, N.et alAutoinducer of virulence as a target for vaccine and therapy against Staphylococcusaureus. Science 280, 438-440 (1995); Balaban, N., Goldkorn, T., Gov, Y., Hirshberg, M., Koyfman, N., Matthews, H.R., Nhan , R.T., Singh, B., and Uziel, O. (2001) J. Biol. ...

Claims

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Application Information

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IPC IPC(8): C07K14/31C12N9/36A61K38/48A61P31/04
Inventor 杨光邵宁生高亚萍董洁李少华柳川沈倍奋范明
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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