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Compound capable of binding to naturally occurring denatured protein, and method for screening for the compound

A screening method, protein technology, applied in the field of compounds combined with natural denatured proteins and the screening thereof

Inactive Publication Date: 2013-01-30
PRISM BIOLAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, screening for irregular sequences has never been looked at

Method used

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  • Compound capable of binding to naturally occurring denatured protein, and method for screening for the compound
  • Compound capable of binding to naturally occurring denatured protein, and method for screening for the compound
  • Compound capable of binding to naturally occurring denatured protein, and method for screening for the compound

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Experimental program
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Effect test

Embodiment 1

[0113] The experimental summary of the published paper (Proc Natl Acad Sci US A. 2004 Aug 24; 101(34): 12682-7) is shown below as a definitive proof of the present invention. Although the inventive idea of ​​the present invention is not disclosed in this paper, the content described in this paper constitutes this specification. The paper only gives that ICG-001 as a peptidomimetic compound only binds to CBP, and further binds to the N-terminal sequence of CBP, but does not bind to p300. However, it is clear that it proves the technical idea of ​​the present invention.

[0114] ICG-001 is shown in Chemical Formula 1, and the biotinylated product of ICG-001 is shown in Chemical Formula 2 (ICG-002). The results of analysis by immunoprecipitation using these substances are shown in figure 1 . Band 1 was the result of incubating the nuclear extract of SW480 cells (a cell line derived from human colon cancer) with the product bound to ICG-002 on streptavidin agarose beads. Band ...

Embodiment 2

[0118] Expressed in Escherichia coli 15 N-tagged CBP(1-111) (indeterminate sequence region) was dissolved in buffer (NaP 25mM·pH6.8, NaCl 50mM, 0.02% NaN 3 ), put 300 μl in the NMR sample tube. Furthermore, adding D of ICG-001 2 O solution brought ICG-001 to concentrations of 5 μM and 15 μM. The solution thus prepared was used as an NMR sample for NMR analysis. Measurement and analysis are carried out in NMR analysis, and a 2D spectrum is obtained in which the X-axis shows the C-13 data and the Y-axis shows the N-15 data.

[0119] The results of CBP1-111 are shown in Figure 4 , the results of CBP1-111 co-existing with 5 μM ICG-001 are shown in Figure 5 , the results of CBP1-111 co-existing with 15 μM ICG-001 are shown in Figure 6 . As a result, CBP1-111 showed peaks typical of native denatured proteins (see Figure 4 ), the change of the peak can be seen by adding ICG-001 (refer to Figure 5-6 ). From this result, a clear interaction was observed between CBP1-111 ...

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Abstract

The screening for a low-molecular-weight compound capable of controlling a protein-protein interaction has been made among from many proteins each having an irregular sequence including a transcription factor. However, it was almost impossible to obtain a low-molecular-weight compound having a satisfactory activity. It is found that the screening for a protein capable of controlling the interaction between a naturally occurring denatured protein and a partner protein can be achieved when attention is focused on irregular sequences of proteins. It is also found that a compound capable of controlling the activity of a naturally occurring denatured protein with high efficiency can be screened by selecting a candidate compound from peptide mimic compounds each having a peptide mimic backbone and a peptide side chain or a side chain analogous to the peptide side chain.

Description

technical field [0001] The present invention relates to a compound that binds to an irregular sequence region (hereinafter sometimes also referred to as an irregular region) of a natural denatured protein. In detail, the present invention relates to a compound that binds to the irregular sequence region of the naturally denatured protein, and utilizes the combination to change the structure of the irregular sequence domain, thereby resulting in an incompatibility with the natural state of the naturally denatured protein. It binds to the cooperating protein that should be bound under it, thereby hindering the biological reaction that occurs in the natural state. In addition, the present invention also relates to screening methods for compounds that bind to irregular sequence regions of natural denatured proteins, and the like. Furthermore, the present invention also relates to medicaments containing compounds that bind to irregular sequence regions of native denatured proteins...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/15G01N33/50
CPCG01N2500/02G01N2500/04G01N33/6872
Inventor 小路弘行
Owner PRISM BIOLAB