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A method for co-crystallization of protein target molecules and small chemical molecules

A small molecule and protein technology, applied in organic chemistry methods, chemical instruments and methods, organic chemistry, etc., can solve problems such as unused crystal screening methods

Active Publication Date: 2021-09-10
徐竞
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  • Application Information

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Problems solved by technology

Until now, this technique of co-crystallization has not been used in crystal screening methods

Method used

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  • A method for co-crystallization of protein target molecules and small chemical molecules
  • A method for co-crystallization of protein target molecules and small chemical molecules
  • A method for co-crystallization of protein target molecules and small chemical molecules

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Embodiment 1

[0026] This example takes the co-crystallization of 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase as an example, and the specific steps are as follows:

[0027] 1) Immobilization of chemical small molecules on the orifice plate: select the chemical small molecule ligands that need to be screened, completely dissolve them in methanol organic solvent to form a solution with a concentration of 1000mmol / L, and divide the solution at a separation factor of 15000g Centrifuge for 15 minutes, take 2 μL of the 500 mmol / L solution that has been centrifuged and remove impurities, and manually drop it into a 24-well plate or a 96-well plate using the convective equilibrium sitting drop method, and then add 0.1 μL of D solution to mix it evenly. Dry in a vacuum drying oven for 24 hours to obtain solidified molecules, wherein the D solution is obtained by mixing the binder, the slow-release agent and the solubilizer with a volume ratio of 1:1:1, and then filtering it; the binder i...

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Abstract

The invention discloses a co-crystallization method of a protein target molecule and a small chemical molecule. The chemical small molecule is dissolved in an organic solvent to form a solution, and after the solution is centrifuged to remove impurities, the supernatant is dropped into an orifice plate, and the Add a solution mixed with a binder, a slow-release agent and a solubilizer, dry and solidify, then add a protein buffer solution containing a precipitating agent and a seed crystal to the dry and solidified molecules, at 4‑20 ° C, neutral The co-crystallization culture is carried out in an environment. This preparation method has the advantages of short time consumption, simple operation and high success rate, and is conducive to automation and high-throughput ligand screening.

Description

technical field [0001] The invention belongs to the technical field of co-crystallization, in particular to protein molecules and small molecules, and in particular to a method for co-crystallization of protein target molecules and chemical small molecules. Background technique [0002] In drug research and development, the three-dimensional spatial structure of proteins and the spatial combination of proteins and active compound molecules are particularly important. It is an indispensable decisive link for drug research and development to transform from traditional random screening to precise and intelligent drug design. Protein crystallography is almost the only practical technique to provide ultra-high resolution of proteins. Today, protein crystallography has been used as one of the routine key technologies and has been most widely used in the world. And high-throughput protein crystallography will remain central to future drug discovery research. Compared with traditi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/30C07C51/43C07C59/265
CPCC07B2200/13C07C51/43C07K1/306C07C59/265
Inventor 傅勘
Owner 徐竞
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