Abstract
Crystal structures of haloalkane dehalogenase were determined in the presence of the substrate 1,2-dichloroethane. At pH 5 and 4 °C, substrate is bound in the active site without being converted; warming to room temperature causes the substrate's carbon–chlorine bond to be broken, producing a chloride ion with concomitant alkylation of the active-site residue Asp124. At pH 6 and room temperature the alkylated enzyme is hydrolysed by a water molecule activated by the His289–Asp260 pair in the active site. These results show that catalysis by the dehalogenase proceeds by a two-step mechanism involving an ester intermediate covalently bound at Asp124.
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Verschueren, K., Seljée, F., Rozeboom, H. et al. Crystallographic analysis of the catalytic mechanism of haloalkane dehalogenase. Nature 363, 693–698 (1993). https://doi.org/10.1038/363693a0
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DOI: https://doi.org/10.1038/363693a0
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