Computational investigation of orotate phosphoribosyltransferase (OPRT) action, an enzymatic reaction between phosphoribosyl pyrophosphate (PRPP) and orotic acid (OA) to yield orotidine 5'-monophosphate (OMP), was carried out. Insights into the pathways of the substrate attack step of the reaction were developed under the quantum mechanics/molecular mechanics framework with S. cerevisiae strain as the representative enzyme bearer. Four pathways were proposed for PRPP and OA binding differing in the sequence of PRPP, OA and Mg²⁺ ion complexation with OPRT. The formation of Mg²⁺-OPRT complex was accompanied by a small energy change while the largest stabilization was observed for the formation of Mg²⁺-PRPP complex supporting the experimental observation of Mg²⁺-PRPP complex as the true substrate for the reaction. Formation of PRPP-OPRT complex was found to be energetically not probable rendering the pathway requiring Mg²⁺-OA complex not probable. Further, PRPP migration towards the active site was found to be energetically not favoured rendering the pathway involving Mg²⁺-OA complexation improbable. Migration of OA and Mg²⁺-PRPP complex towards the active site was found to be energetically probable with a large stabilization of the system when Mg²⁺-PRPP complex bound to the OA-OPRT complex. This conclusively proved the sequential binding of OA and Mg²⁺-PRPP complexes during OPRT action.

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Journal Article

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Subrahmanyeswara Rao NN, Deshpande PA

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