Stabilization of Different Types of Transition States in a Single Enzyme Active Site: QM/MM Analysis of Enzymes in the Alkaline Phosphatase Superfamily.

The first step for the hydrolysis of a phosphate monoester (pNPP) in enzymes of the alkaline phosphatase (AP) superfamily, R166S AP and wild-type NPP, is studied using QM/MM simulations based on an approximate density functional theory (SCC-DFTBPR) and a recently introduced QM/MM interaction Hamilto...

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Publicado en:Journal of the American Chemical Society Vol. 135; no. 28; pp. 10457 - 10470
Autores principales: Guanhua Hou, Qiang Cui
Formato: Artículo
Publicado: American Chemical Society 7/17/2013
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Acceso en línea:Ver este registro en EBSCOhost
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      pub: American Chemical Society
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        10.1021/ja403293d
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        atl: Stabilization of Different Types of Transition States in a Single Enzyme Active Site: QM/MM Analysis of Enzymes in the Alkaline Phosphatase Superfamily.
      aug:
        au:
          Guanhua Hou
          Qiang Cui
        affil: Department of Chemistry and Theoretical Chemistry Institute, University of Wisconsin-Madison, 1101 University Avenue, Madison, Wisconsin 53706, United States
      su:
        Transition state theory (Chemistry)
        Enzymes
        Alkaline phosphatase
        Hydrolysis
        Density functional theory
        Phosphates
        Substrates (Materials science)
      sug:
        subj:
          Transition state theory (Chemistry)
          Enzymes
          Alkaline phosphatase
          Hydrolysis
          Density functional theory
          Phosphates
          Substrates (Materials science)
      ab: The first step for the hydrolysis of a phosphate monoester (pNPP) in enzymes of the alkaline phosphatase (AP) superfamily, R166S AP and wild-type NPP, is studied using QM/MM simulations based on an approximate density functional theory (SCC-DFTBPR) and a recently introduced QM/MM interaction Hamiltonian. The calculations suggest that similar loose transition states are involved in both enzymes, despite the fact that phosphate monoesters are the cognate substrates for AP but promiscuous substrates for NPP. The computed loose transition states are clearly different from the more synchronous ones previously calculated for diester reactions in the same AP enzymes. Therefore, our results explicitly support the proposal that AP enzymes are able to recognize and stabilize different types of transition states in a single active site. Analysis of the structural features of computed transition states indicates that the plastic nature of the bimetallic site plays a minor role in accommodating multiple types of transition states and that the high degree of solvent accessibility of the AP active site also contributes to its ability to stabilize diverse transition-state structures without the need of causing large structural distortions of the bimetallic motif. The binding mode of the leaving group in the transition state highlights that vanadate may not always be an ideal transition state analog for loose phosphoryl transfer transition states.
      pubtype: Academic Journal
      doctype: Article
      src: R
    language: English
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