Research Article
Examining the promiscuous phosphatase activity of Pseudomonas aeruginosa arylsulfatase: A comparison to analogous phosphatases
Jinghui Luo,
Bert van Loo,
S. C. L. Kamerlin,
Jinghui Luo
Department of Cell and Molecular Biology (ICM), Uppsala University, Uppsala Biomedical Center (BMC), S-75124 Uppsala, Sweden
Department of Biophysical and Structural Chemistry, Leiden Institute of Chemistry, Leiden University, 2300 RA Leiden, The Netherlands
Search for more papers by this authorBert van Loo
Department of Biochemistry, University of Cambridge, CB2 1GA Cambridge, United Kingdom
Search for more papers by this authorCorresponding Author
S. C. L. Kamerlin
Department of Cell and Molecular Biology (ICM), Uppsala University, Uppsala Biomedical Center (BMC), S-75124 Uppsala, Sweden
Department of Cell and Molecular Biology (ICM), Uppsala University, Uppsala Biomedical Center (BMC), S-75124 Uppsala, Sweden===Search for more papers by this authorJinghui Luo,
Bert van Loo,
S. C. L. Kamerlin,
Jinghui Luo
Department of Cell and Molecular Biology (ICM), Uppsala University, Uppsala Biomedical Center (BMC), S-75124 Uppsala, Sweden
Department of Biophysical and Structural Chemistry, Leiden Institute of Chemistry, Leiden University, 2300 RA Leiden, The Netherlands
Search for more papers by this authorBert van Loo
Department of Biochemistry, University of Cambridge, CB2 1GA Cambridge, United Kingdom
Search for more papers by this authorCorresponding Author
S. C. L. Kamerlin
Department of Cell and Molecular Biology (ICM), Uppsala University, Uppsala Biomedical Center (BMC), S-75124 Uppsala, Sweden
Department of Cell and Molecular Biology (ICM), Uppsala University, Uppsala Biomedical Center (BMC), S-75124 Uppsala, Sweden===Search for more papers by this authorAbstract
Pseudomonas aeruginosa arylsulfatase (PAS) is a bacterial sulfatase capable of hydrolyzing a range of sulfate esters. Recently, it has been demonstrated to also show very high proficiency for phosphate ester hydrolysis. Such proficient catalytic promiscuity is significant, as promiscuity has been suggested to play an important role in enzyme evolution. Additionally, a comparative study of the hydrolyses of the p-nitrophenyl phosphate and sulfate monoesters in aqueous solution has demonstrated that despite superficial similarities, the two reactions proceed through markedly different transition states with very different solvation effects, indicating that the requirements for the efficient catalysis of the two reactions by an enzyme will also be very different (and yet they are both catalyzed by the same active site). This work explores the promiscuous phosphomonoesterase activity of PAS. Specifically, we have investigated the identity of the most likely base for the initial activation of the unusual formylglycine hydrate nucleophile (which is common to many sulfatases), and demonstrate that a concerted substrate-as-base mechanism is fully consistent with the experimentally observed data. This is very similar to other related systems, and suggests that, as far as the phosphomonoesterase activity of PAS is concerned, the sulfatase behaves like a “classical” phosphatase, despite the fact that such a mechanism is unlikely to be available to the native substrate (based on pKa considerations and studies of model systems). Understanding such catalytic versatility can be used to design novel artificial enzymes that are far more proficient than the current generation of designer enzymes. Proteins 2012; © 2011 Wiley Periodicals, Inc.
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