Hit expansion for ligand 4.

<p>For chemical structures see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035792#pone-0035792-g013" target="_blank">Figure 13</a>.</p>*<p>average values of three independent measurements, standard deviation in brackets.</p>a<p>no inhibition at solubility limit measured.</p

Hit expansion for ligand 8.

<p>For chemical structures see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035792#pone-0035792-g016" target="_blank">Figure 16</a>.</p>*<p>average values of three independent measurements, standard deviation in brackets.</p>a<p>no inhibition at solubility limit measured.</p

Docking ranks, physico-chemical properties, inhibition values, and ligand efficiencies for virtual screening hits.

<p>For chemical structures see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035792#pone-0035792-g007" target="_blank">Figure 7</a>.</p>1<p>using the total score of docking for ranking (after application of the pharmacophore filter).</p>2<p>using the score divided by the number of heavy atoms of the molecule for ranking (after application of the pharmacophore filter).</p>3<p>setup 1: His25 protonated at ND, no ADP present; setup 2: His25 protonated at ND, ADP present; setup 3: His25 protonated at NE, no ADP present; setup 4: His25 protonated at NE, ADP present.</p>4<p>average values of three independent measurements, standard deviation in brackets.</p

Physico-chemical properties, inhibition values, and ligand efficiencies for <i>in vitro</i> screening hits.

<p>For chemical structures see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035792#pone-0035792-g009" target="_blank">Figure 9</a>.</p>*<p>average values of three independent measurements, standard deviation in brackets.</p

Chemical structures of <i>in vitro</i> screening hits.

<p>Chemical structures of <i>in vitro</i> screening hits.</p

Substrate binding site of <i>Aa</i>IspE (PDB code 2v2z).

<p>The catalytic residues Lys9 and Asp130 are labelled together with other residues important for ligand binding. The cytidine moiety of the substrate forms hydrogen bonds with Lys145 and His25, π-stacking interactions with Tyr175 and edge-face interactions with Tyr24.</p

Superposition of adenine binding sites of human CDK2 and <i>Aa</i>IspE.

<p>ATP adopts the <i>anti</i> conformation in <i>h</i>CDK2 (PDB code 2cch, carbon atoms coloured pink) while the ATP analogue AMP-PNP adopts the <i>syn</i> conformation in <i>Aa</i>IspE (PDB code 2v8p, carbon atoms coloured light blue). Further, in <i>h</i>CDK2 adenine forms hydrogen bonds with the backbone amide groups of the hinge region while in <i>Aa</i>IspE the backbone and side chain atoms of surrounding amino acids are involved in hydrogen bonding-interactions. In both enzymes, a donor-acceptor-donor motive (green circle) is important for molecular recognition.</p

Hit expansion for ligand 3.

<p>For chemical structures see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035792#pone-0035792-g011" target="_blank">Figure 11</a>.</p>*<p>average values of three independent measurements, standard deviation in brackets.</p

Chemical structures for analogues of ligand 4.

<p>Chemical structures for analogues of ligand 4.</p

Ranks for the <i>in silico</i> and <i>in vitro</i> screening hits when the HTS library is docked against <i>Aa</i>IspE without prior filtering for physico-chemical properties or pharmacophore constraints.

<p>For chemical structures see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035792#pone-0035792-g007" target="_blank">Figure 7</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035792#pone-0035792-g009" target="_blank">Figure 9</a>.</p>1<p>using the total score of docking for ranking.</p>2<p>using the score divided by the number of heavy atoms of the molecule for ranking.</p>3<p>setup 1: His25 protonated at ND, no ADP present; setup 2: His25 protonated at ND, ADP present; setup 3: His25 protonated at NE, no ADP present; setup 4: His25 protonated at NE, ADP present.</p