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The crystal structure of phenylpyruvate decarboxylase from Azospirillum brasilense at 1.5 Å resolution
Author(s) -
Versées Wim,
Spaepen Stijn,
Vanderleyden Jos,
Steyaert Jan
Publication year - 2007
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2007.05771.x
Subject(s) - allosteric regulation , biochemistry , cofactor , active site , pyruvate decarboxylase , decarboxylation , azospirillum brasilense , biology , chemistry , stereochemistry , enzyme , bacteria , catalysis , alcohol dehydrogenase , genetics , nitrogen fixation
Phenylpyruvate decarboxylase (PPDC) of Azospirillum brasilense , involved in the biosynthesis of the plant hormone indole‐3‐acetic acid and the antimicrobial compound phenylacetic acid, is a thiamine diphosphate‐dependent enzyme that catalyses the nonoxidative decarboxylation of indole‐ and phenylpyruvate. Analogous to yeast pyruvate decarboxylases, PPDC is subject to allosteric substrate activation, showing sigmoidal v versus [S] plots. The present paper reports the crystal structure of this enzyme determined at 1.5 Å resolution. The subunit architecture of PPDC is characteristic for other members of the pyruvate oxidase family, with each subunit consisting of three domains with an open α/β topology. An active site loop, bearing the catalytic residues His112 and His113, could not be modelled due to flexibility. The biological tetramer is best described as an asymmetric dimer of dimers. A cysteine residue that has been suggested as the site for regulatory substrate binding in yeast pyruvate decarboxylase is not conserved, requiring a different mechanism for allosteric substrate activation in PPDC. Only minor changes occur in the interactions with the cofactors, thiamine diphosphate and Mg 2+ , compared to pyruvate decarboxylase. A greater diversity is observed in the substrate binding pocket accounting for the difference in substrate specificity. Moreover, a catalytically important glutamate residue conserved in nearly all decarboxylases is replaced by a leucine in PPDC. The consequences of these differences in terms of the catalytic and regulatory mechanism of PPDC are discussed.