In silico analysis of RB and hepatocyte growth factor receptor (MET) mutant proteins
The p.(Gln850Ter) mutation of RB protein causes a truncation of the full-length protein (1-928) by 78 residues (Figure 3 ). The deletion of the C-terminal region (851-928 residues) was predicted to cause no alteration of the secondary structural arrangement, neither local nor long-distance effect were predicted for the truncated protein. Disorder prediction also showed that the truncation of the protein does not cause significant changes in pathogenicity.
Neither secondary structure nor disorder predictions showed any adverse effects of the deletion on protein structure. Therefore, the consequence of the truncation can be associated with the loss of functional regions. The functional importance of the C-terminal region in RB protein is implied by the presence of ELMs, including a bipartite nuclear localization signal (860-876). Consequently, the loss of this signal sequence may result an impaired nuclear localization of the mutant/truncated protein, while the wild-type protein naturally enters the nucleus. The structure and possible interactions of RB1 protein was presented in Figure 3 . The following coordinate files were used to prepare the figure: 2QDJ.pdb [14]; 4ELL.pdb [15]; 2AZE.pdb [16]; 3N5U.pdb [17]; 1H25.pdb [18]; 1PJM [19].
The p.(Thr1010Ile) mutation of MET protein causes a non-synonymous mutation of a polar threonine to a hydrophobic isoleucine residue. The possible effects of this point mutation were predicted by multiple algorithms which showed no alteration of the secondary structural arrangement at or in the proximity of the mutated residue, the disorder propensity predicted for the mutant were highly similar to those obtained for the wild-type. In agreement with this, the sequence-based prediction of stability changes also implied neutral nature of the mutation (-0.37 kcal/mol). We predicted no significant increase or decrease of free energy change value upon p.(Thr992Ile) mutation. Similar to RB protein, no local or global disturbances of the structure could be predicted, rather functional changes of the residue in 992th position may be responsible for the phenotypic effects.