Ure S7A). The total percentage of patients with c.38G.

Ure S7A). The total percentage of patients with c.38G.A (p.G13D) MedChemExpress Conduritol B epoxide mutations was 18.6 (Figure S7B) and 8.73 when considering the entire CRC population (Figure S7A). However, this frequency may differ between populations; forFigure 4. Distributions of docking scores. The docking scores for WT (blue), G12D (red) and G13D (green) KRAS proteins docked with GTP are shown in different colors. doi:10.1371/journal.pone.0055793.gComputational Analysis of KRAS MutationsFigure 5. Free energy profiles determined for the binding of GTP with WT (blue), G12D (red) and G13D (green). The reaction coordinate is defined as the distance between the mass center of the GTP and the mass center of the WT, G12D, and G13D KRAS proteins, respectively. doi:10.1371/journal.pone.0055793.order GDC-0917 ginstance, in the Taiwanese population, this frequency ranged from 33 to 40 [52,53]. Such population differences could be because of detection limits of the methodologies used and the homogeneity of specimens. However, taking together different international published series, mutations in c.35G.A (p.G12D) and c.35G.T (p.G12V) were reported to be the most frequent and c.34G.A (p.G12S), c.35G.C (p.G12R) and c.34G.C (p.G12R) were reported to be the least frequent [54,55]. Moreover, mutations in codon 13, in which the glycine changes to aspartate, account for over 80 of the mutations occurring in this codon and approximately 19 of KRAS mutations in patients [55,56]. Similarly, the prevalence of KRAS mutations in codon 12 and 13 was 79 and 21 , respectively, in the Taiwanese population (unpublished data, total 420 CRC patients). Because the mutations in KRAS codon 13 represent a high proportion of the population suffering from CRC, the issue regarding the possible response of this mutated form of KRAS to anti-EGFR therapies should be considered relevant from biological, ethical and economical perspectives. Different KRAS mutations in codon 12 could be associated with distinctive clinicopathological feature. Clinical studies shows that the KRAS codon 12 mutation, especially the c.35G.T (p.G12V) mutation, was associated with the highest colorectal cancer?specific mortality [25]. However, an experimental study showed that c.34G.C (p.G12R) and c.35G.T (p.G12V) mutations confer more potent transforming ability than other KRAS mutations, including c.34G.A (p.G12S), c.34G.T (p.G12C), c.35G.A (p.G12D), and c.35G.C (p.G12A) [17]. Furthermore, the GTPase activity of c.34G.C (p.G12R) and c.35G.T (p.G12V) mutants is lower than that of other KRAS mutations [57,58]. Previously, MD calculations were performed on the wildtype p21ras, the p.G12V oncogenic mutant p21ras, and the p.G12P non-oncogenic mutant p21ras [59]. The results indicated that the structure of the active site of the enzyme substrate complex in the oncogenic mutant p21ras continuously changes, and these changes in the active site would make it difficult for the GTPGDP hydrolysis reaction 23388095 to occur in the mutant. Recently, Messner et al. [60] indicated that p.G13D mutated CRC cells are more sensitive to anti-EGFR treatment than codon 12-or codon 61mutated cells and the p.G13D-mutated CRC cells seem to define a less aggressive phenotype. Similarly, De Roock W et al. [26] suggested that p.G12V-mutated cells were insensitive to cetuximab, however, p.G13-mutated cells were nearly as response to cetuximab as wild-type cells. The rate of GTP-to-GDP conversion can be dramatically accelerated by an accessory protein of the guanine nucleotide a.Ure S7A). The total percentage of patients with c.38G.A (p.G13D) mutations was 18.6 (Figure S7B) and 8.73 when considering the entire CRC population (Figure S7A). However, this frequency may differ between populations; forFigure 4. Distributions of docking scores. The docking scores for WT (blue), G12D (red) and G13D (green) KRAS proteins docked with GTP are shown in different colors. doi:10.1371/journal.pone.0055793.gComputational Analysis of KRAS MutationsFigure 5. Free energy profiles determined for the binding of GTP with WT (blue), G12D (red) and G13D (green). The reaction coordinate is defined as the distance between the mass center of the GTP and the mass center of the WT, G12D, and G13D KRAS proteins, respectively. doi:10.1371/journal.pone.0055793.ginstance, in the Taiwanese population, this frequency ranged from 33 to 40 [52,53]. Such population differences could be because of detection limits of the methodologies used and the homogeneity of specimens. However, taking together different international published series, mutations in c.35G.A (p.G12D) and c.35G.T (p.G12V) were reported to be the most frequent and c.34G.A (p.G12S), c.35G.C (p.G12R) and c.34G.C (p.G12R) were reported to be the least frequent [54,55]. Moreover, mutations in codon 13, in which the glycine changes to aspartate, account for over 80 of the mutations occurring in this codon and approximately 19 of KRAS mutations in patients [55,56]. Similarly, the prevalence of KRAS mutations in codon 12 and 13 was 79 and 21 , respectively, in the Taiwanese population (unpublished data, total 420 CRC patients). Because the mutations in KRAS codon 13 represent a high proportion of the population suffering from CRC, the issue regarding the possible response of this mutated form of KRAS to anti-EGFR therapies should be considered relevant from biological, ethical and economical perspectives. Different KRAS mutations in codon 12 could be associated with distinctive clinicopathological feature. Clinical studies shows that the KRAS codon 12 mutation, especially the c.35G.T (p.G12V) mutation, was associated with the highest colorectal cancer?specific mortality [25]. However, an experimental study showed that c.34G.C (p.G12R) and c.35G.T (p.G12V) mutations confer more potent transforming ability than other KRAS mutations, including c.34G.A (p.G12S), c.34G.T (p.G12C), c.35G.A (p.G12D), and c.35G.C (p.G12A) [17]. Furthermore, the GTPase activity of c.34G.C (p.G12R) and c.35G.T (p.G12V) mutants is lower than that of other KRAS mutations [57,58]. Previously, MD calculations were performed on the wildtype p21ras, the p.G12V oncogenic mutant p21ras, and the p.G12P non-oncogenic mutant p21ras [59]. The results indicated that the structure of the active site of the enzyme substrate complex in the oncogenic mutant p21ras continuously changes, and these changes in the active site would make it difficult for the GTPGDP hydrolysis reaction 23388095 to occur in the mutant. Recently, Messner et al. [60] indicated that p.G13D mutated CRC cells are more sensitive to anti-EGFR treatment than codon 12-or codon 61mutated cells and the p.G13D-mutated CRC cells seem to define a less aggressive phenotype. Similarly, De Roock W et al. [26] suggested that p.G12V-mutated cells were insensitive to cetuximab, however, p.G13-mutated cells were nearly as response to cetuximab as wild-type cells. The rate of GTP-to-GDP conversion can be dramatically accelerated by an accessory protein of the guanine nucleotide a.