For the calculation strategy of CASTp, alpha-shape and discrete-flow methods are applied to the protein binding site, also the measurement of pocket size by the program of CAST by Liang et al. in 1998, then updated by Tian et al. in 2018. Firstly, CAST identifies atoms which form the protein pocket, then calculates the volume and area, identifies the atoms forming the rims of pocket mouth, computes how many mouth openings for each pocket, predict the area and circumference of mouth openings, finally locates cavities and calculate their size. The secondary structures were calculated by DSSP. The single amino acid annotations were fetched from UniProt database, then mapped to PDB structures following residue-level information from SIFTS database.