Each structure is used as a design scaffold for each mutation

Each get Fingolimod (hydrochloride) structure is used as a design scaffold for each mutation). Test cases are: CA: Carbonic anhydrase II, ABP D7r4 amine binding protein, ER: Estrogen receptor a, HP: HIV-1 protease, KI: Ketosteroid isomerase, L: Lectin, MS: Methylglyoxal synthase, N1: Neuroaminidase test 1, N2: Neuroaminidase test 2, PNP: Purine nucleoside phosphorylase, S1: Streptavidin test 1, S2: Streptavidin test 2, TS: Thymidylate synthase, T: Trypsin. doi:10.1371/journal.pone.0052505.gTable 2. Order of designs by predicted binding score.Test CasesCADDSuite Total Binding 1/1 1/1 6/9 2/2 0/2 K 6/8 4/4 2/2 0/6 K 65.6Vina Total 1/1 1/1 9/9 2/2 0/2 1/2 7/8 4/4 2/2 1/6 1/2 75.5 Binding 1/1 1/1 9/9 2/2 0/2 0/2 6/8 4/4 2/2 0/6 1/2 68.8Rosetta Total 1/1 1/1 5/9 1/2 1/2 0/2 4/8 3/4 2/2 6/6 0/2 63.5 Binding 1/1 1/1 8/9 1/2 0/2 2/2 2/8 3/4 1/2 3/6 1/2 63.5D7r4 amine binding protein Estrogen receptor HIV-1 protease Ketosteroid isomerase Neuroaminidase test 1 Neuroaminidase test 2 Purine nucleoside phosphorylase Streptavidin test 1 Streptavidin test 2 Thymidylate synthase Trypsin Mean1/1 1/1 6/9 2/2 0/2 2/2 6/8 4/4 2/2 1/6 1/2 70.8Numbers of correctly ranked design mutation pairs with large affinity difference. All mutation pairs for which there is an affinity difference of at least 50-fold are investigated. All design pairs with these mutations (i.e. for each of these pairs there are as many design pairs as scaffold crystal structures) are checked, if the order of the mutations by total score or binding score is the same order as by affinity. A cell shows the number of correctly ordered design pairs, and the number of all design pairs. The mean for this part is calculated by scaling the percentage of a test case by the number of mutation pairs (i.e. NOT by design pairs, which would bias the value too much towards test cases with many crystal structures). doi:10.1371/journal.pone.0052505.tComputational Design of Binding PocketsFigure 4. Comparison of the energy scores versus the affinities of the mutations show how well the programs reproduce the differences. For each test case with more than two mutations, we plotted the top binding scores of CADDSuite, Vina, and Rosetta designs for each mutation on each scaffold structure together with the logarithm of the affinity. Here we 24195657 show plots for Carbonic anhydrase II, HIV-1 protease, and Streptavidin test 1. All other plots are shown in Information S1. Values are FG-4592 site scaled to fit in the same range. Shown on the x-axis of a plot are the mutants in order of affinity to the ligand (the leftmost has the lowest affinity, compare Table 1 for the actual values). The y-axis measures predicted binding scores for the designs, and the log affinities, scaled between 0 and 1. Both are proportional to the binding free energy, and can therefore be compared when scaled to the same range. The lowest predicted binding score or log affinity is set to 0, the highest respective value to 1. Each plot contains a line for the affinity logarithm (solid, black no marker). This line represents the goal, if a method predicts binding well, the binding score lines should closely follow the log affinity line. The other markers and lines show the scaled predicted binding scores. One line represents the designs calculated for all available mutants, calculated by using one crystal structure as the scaffold. (Crystal structure 1: dashed, blue, circle markers; structure 2: red, dotted, square markers; structure 3: green, dash-dot pattern, diamond markers; structure.Each structure is used as a design scaffold for each mutation). Test cases are: CA: Carbonic anhydrase II, ABP D7r4 amine binding protein, ER: Estrogen receptor a, HP: HIV-1 protease, KI: Ketosteroid isomerase, L: Lectin, MS: Methylglyoxal synthase, N1: Neuroaminidase test 1, N2: Neuroaminidase test 2, PNP: Purine nucleoside phosphorylase, S1: Streptavidin test 1, S2: Streptavidin test 2, TS: Thymidylate synthase, T: Trypsin. doi:10.1371/journal.pone.0052505.gTable 2. Order of designs by predicted binding score.Test CasesCADDSuite Total Binding 1/1 1/1 6/9 2/2 0/2 K 6/8 4/4 2/2 0/6 K 65.6Vina Total 1/1 1/1 9/9 2/2 0/2 1/2 7/8 4/4 2/2 1/6 1/2 75.5 Binding 1/1 1/1 9/9 2/2 0/2 0/2 6/8 4/4 2/2 0/6 1/2 68.8Rosetta Total 1/1 1/1 5/9 1/2 1/2 0/2 4/8 3/4 2/2 6/6 0/2 63.5 Binding 1/1 1/1 8/9 1/2 0/2 2/2 2/8 3/4 1/2 3/6 1/2 63.5D7r4 amine binding protein Estrogen receptor HIV-1 protease Ketosteroid isomerase Neuroaminidase test 1 Neuroaminidase test 2 Purine nucleoside phosphorylase Streptavidin test 1 Streptavidin test 2 Thymidylate synthase Trypsin Mean1/1 1/1 6/9 2/2 0/2 2/2 6/8 4/4 2/2 1/6 1/2 70.8Numbers of correctly ranked design mutation pairs with large affinity difference. All mutation pairs for which there is an affinity difference of at least 50-fold are investigated. All design pairs with these mutations (i.e. for each of these pairs there are as many design pairs as scaffold crystal structures) are checked, if the order of the mutations by total score or binding score is the same order as by affinity. A cell shows the number of correctly ordered design pairs, and the number of all design pairs. The mean for this part is calculated by scaling the percentage of a test case by the number of mutation pairs (i.e. NOT by design pairs, which would bias the value too much towards test cases with many crystal structures). doi:10.1371/journal.pone.0052505.tComputational Design of Binding PocketsFigure 4. Comparison of the energy scores versus the affinities of the mutations show how well the programs reproduce the differences. For each test case with more than two mutations, we plotted the top binding scores of CADDSuite, Vina, and Rosetta designs for each mutation on each scaffold structure together with the logarithm of the affinity. Here we 24195657 show plots for Carbonic anhydrase II, HIV-1 protease, and Streptavidin test 1. All other plots are shown in Information S1. Values are scaled to fit in the same range. Shown on the x-axis of a plot are the mutants in order of affinity to the ligand (the leftmost has the lowest affinity, compare Table 1 for the actual values). The y-axis measures predicted binding scores for the designs, and the log affinities, scaled between 0 and 1. Both are proportional to the binding free energy, and can therefore be compared when scaled to the same range. The lowest predicted binding score or log affinity is set to 0, the highest respective value to 1. Each plot contains a line for the affinity logarithm (solid, black no marker). This line represents the goal, if a method predicts binding well, the binding score lines should closely follow the log affinity line. The other markers and lines show the scaled predicted binding scores. One line represents the designs calculated for all available mutants, calculated by using one crystal structure as the scaffold. (Crystal structure 1: dashed, blue, circle markers; structure 2: red, dotted, square markers; structure 3: green, dash-dot pattern, diamond markers; structure.