Structural characterization of 2,6-dichloro-p-hydroquinone 1,2-dioxygenase (PcpA) from Sphingobium chlorophenolicum, a new type of aromatic ring-cleavage enzyme

Robert P Hayes; Abigail R Green; Mark S Nissen; Kevin M Lewis; Luying Xun; ChulHee Kang

doi:10.1111/mmi.12204

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Structural characterization of 2,6-dichloro-p-hydroquinone 1,2-dioxygenase (PcpA) from Sphingobium chlorophenolicum, a new type of aromatic ring-cleavage enzyme

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Structural characterization of 2,6-dichloro-p-hydroquinone 1,2-dioxygenase (PcpA) from Sphingobium chlorophenolicum, a new type of aromatic ring-cleavage enzyme

Robert P Hayes, Abigail R Green, Mark S Nissen, Kevin M Lewis, Luying Xun and ChulHee Kang

Molecular microbiology, Vol.88(3), pp.523-536

05/2013

DOI: https://doi.org/10.1111/mmi.12204

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https://hdl.handle.net/2376/115437

PMCID: PMC4211620

PMID: 23489289

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Abstract

PcpA (2,6-dichloro- p -hydroquinone 1,2-dioxygenase) from Sphingobium chlorophenolicum , a non-haem Fe(II) dioxygenase capable of cleaving the aromatic ring of p -hydroquinone and its substituted variants, is a member of the recently discovered p -hydroquinone 1,2-dioxygenases. Here we report the 2.6 Å structure of PcpA, which consists of four βαβββ motifs, a hallmark of the vicinal oxygen chelate superfamily. The secondary co-ordination sphere of the Fe(II) centre forms an extensive hydrogen-bonding network with three solvent exposed residues, linking the catalytic Fe(II) to solvent. A tight hydrophobic pocket provides p -hydroquinones access to the Fe(II) centre. The p -hydroxyl group is essential for the substrate-binding, thus phenols and catechols, lacking a p -hydroxyl group, do not bind to PcpA. Site-directed mutagenesis and kinetic analysis confirm the critical catalytic role played by the highly conserved His10, Thr13, His226 and Arg259. Based on these results, we propose a general reaction mechanism for p -hydroquinone 1,2-dioxygenases.

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Title: Structural characterization of 2,6-dichloro-p-hydroquinone 1,2-dioxygenase (PcpA) from Sphingobium chlorophenolicum, a new type of aromatic ring-cleavage enzyme
Creators: Robert P Hayes - Department of Chemistry, Washington State University, Pullman, WA 99164-4630, USA
Abigail R Green - School of Molecular Biosciences, Washington State University, Pullman, WA 99164-4660, USA
Mark S Nissen - Department of Chemistry, Washington State University, Pullman, WA 99164-4630, USA
Kevin M Lewis - Department of Chemistry, Washington State University, Pullman, WA 99164-4630, USA
Luying Xun - School of Molecular Biosciences, Washington State University, Pullman, WA 99164-4660, USA
ChulHee Kang - Department of Chemistry, Washington State University, Pullman, WA 99164-4630, USA
Publication Details: Molecular microbiology, Vol.88(3), pp.523-536
Academic Unit: Chemistry, Department of; Molecular Biosciences, School of
Identifiers: 99900548136501842
Language: English
Resource Type: Journal article