Protein skeleton design method and application thereof
A design method and protein technology, applied in the biological field, can solve the problems of low efficiency, uneven quality of protein structure models, a large number of manual inspections, etc., and achieve the effect of high efficiency and high sequence designability
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Embodiment 1
[0046] Example 1 Design of Small Protein Skeleton and Its Amino Acid Sequence Using Improved Method
[0047] 1) Prepare a small protein-specific HLH fragment database
[0048] In order to improve the HLH fragment assembly efficiency of the SEWING calculation method, the database information in the original method has been adjusted. The original database uses the length of each helix in the HLH fragment to be 5-40 amino acids, which is not suitable for designing small proteins. skeleton. The improved method is to extract non-redundant data with resolution accuracy below 3.0 Å and 30% sequence similarity from the PDB database as the input set of data, and separate HLH fragments with each helical length of 5-25 amino acids. A new database was prepared to control the size of the helical structure within the range of 30-90 amino acids.
[0049] 2) Use the improved SEWING method to generate small protein backbones
[0050] The HLH fragment structure assembled in the SEWING genera...
Embodiment 2
[0066] Example 2 Using a stable small protein backbone structure to design targeted binding drugs
[0067] 1) Isolation of key interacting structural motif fragments from crystal structures.
[0068] Step 1.1: Search the structure of the target protein in the PDB database. If there is a high-precision crystal structure, download its PDB file, and remove irrelevant molecular information such as water molecules, ions, substrates, etc., using the default Rosetta Relax program parameters to optimize the protein structure.
[0069] Step 1.2: Use the Rosetta ddG Mover program to calculate the binding free energy change of the crystal complex structure and scan all residue sites on the interaction interface, and set the amino acids whose free energy contribution is greater than 0.5 REU energy units as hotspot residues . The continuous secondary structures connecting these hotspot residues and the structure of the target were isolated and saved as a new PDB structure.
[0070] 2) M...
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