Protein-Derived Cofactors

Davidson, Victor L.

doi:10.1016/b978-0-12-409547-2.14615-3

“…Cu(II) aids in formation of TPQ in a CuAO-autocatalytic mechanism shown in Figure 5D ( Schwartz et al, 2000 ; Kim et al, 2002 ; Dubois and Klinman, 2005 ; Davidson, 2011 , 2020 ; Klinman and Bonnot, 2014 ). TPQ biosynthesis is a multi-step process, which begins with Cu(II) bound in the active site.…”

Section: Mechanistic Basis Of Plant Copper Metalloenzymesmentioning

confidence: 99%

Plant Copper Metalloenzymes As Prospects for New Metabolism Involving Aromatic Compounds

Mydy

¹

,

Chigumba

²

,

Kersten

³

2021

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Copper is an important transition metal cofactor in plant metabolism, which enables diverse biocatalysis in aerobic environments. Multiple classes of plant metalloenzymes evolved and underwent genetic expansions during the evolution of terrestrial plants and, to date, several representatives of these copper enzyme classes have characterized mechanisms. In this review, we give an updated overview of chemistry, structure, mechanism, function and phylogenetic distribution of plant copper metalloenzymes with an emphasis on biosynthesis of aromatic compounds such as phenylpropanoids (lignin, lignan, flavonoids) and cyclic peptides with macrocyclizations via aromatic amino acids. We also review a recent addition to plant copper enzymology in a copper-dependent peptide cyclase called the BURP domain. Given growing plant genetic resources, a large pool of copper biocatalysts remains to be characterized from plants as plant genomes contain on average more than 70 copper enzyme genes. A major challenge in characterization of copper biocatalysts from plant genomes is the identification of endogenous substrates and catalyzed reactions. We highlight some recent and future trends in filling these knowledge gaps in plant metabolism and the potential for genomic discovery of copper-based enzymology from plants.

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“…The enzyme CD630_32430 (PrdH) catalyzes pyruvoyl cofactor formation in PrdA and is the main focus of this study. C) Proposed mechanism of pyruvoyl cofactor formation in PrdA, based on observations made for pyruvoyl-dependent decarboxylases ( 7 ).…”

Section: Introductionmentioning

confidence: 99%

Formation of the pyruvoyl-dependent proline reductase Prd from Clostridioides difficile requires the maturation enzyme PrdH

Behlendorf,

Diwo,

Neumann-Schaal

et al. 2024

PNAS Nexus

0

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Stickland fermentation, the coupled oxidation and reduction of amino acid pairs, is a major pathway for obtaining energy in the nosocomial bacterium Clostridioides difficile. D-proline is the preferred substrate for the reductive path, making it not only a key component of the general metabolism but also impacting on the expression of the clostridial toxins TcdA and TcdB. D-proline reduction is catalyzed by the proline reductase Prd, which belongs to the pyruvoyl-dependent enzymes. These enzymes are translated as inactive proenzymes and require subsequent processing to install the covalently bound pyruvate. Whereas pyruvoyl formation by intramolecular serinolysis has been studied in unrelated enzymes, details about pyruvoyl generation by cysteinolysis as in Prd are lacking. Here, we show that Prd maturation requires a small dimeric protein that we have named PrdH. PrdH (CD630_32430) is co-encoded with the PrdA and PrdB subunits of Prd and also found in species producing similar reductases. By producing stable variants of PrdA and PrdB, we demonstrate that PrdH-mediated cleavage and pyruvoyl formation in the PrdA subunit requires PrdB, which can be harnessed to produce active recombinant Prd for subsequent analyses. We further created PrdA- and PrdH-mutants to get insight into the interaction of the components and into the processing reaction itself. Finally, we show that deletion of prdH renders C. difficile insensitive to proline concentrations in culture media, suggesting that this processing factor is essential for proline utilization. Due to the link between Stickland fermentation and pathogenesis, we suggest PrdH may be an attractive target for drug development.

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“…PrdF encodes a proline racemase, catalyzing the conversion of L-proline to D-proline. (C) Proposed mechanism of pyruvoyl cofactor formation in PrdA, based on observations made for pyruvoyl-dependent decarboxylases (34).…”

Section: Introductionmentioning

confidence: 99%

Formation of the pyruvoyl-dependent proline reductase Prd fromClostridioides difficilerequires the maturation enzyme PrdH

Behlendorf,

Diwo,

Neumann-Schaal

et al. 2023

Preprint

0

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Stickland fermentation, the coupled oxidation and reduction of amino acid pairs, is a major pathway for obtaining energy in the nosocomial bacteriumClostridioides difficile. D-proline is the preferred substrate for the reductive path, making it not only a key component of the general metabolism but also impacting on the expression of the clostridial toxins TcdA and TcdB. D-proline reduction is catalyzed by the proline reductase Prd, which belongs to the pyruvoyl-dependent enzymes. These enzymes are translated as inactive proenzymes and require subsequent processing to install the covalently bound pyruvate. Whereas pyruvoyl formation by intramolecular serinolysis has been studied in unrelated enzymes, details about pyruvoyl generation by cysteinolysis such as in Prd are lacking. Here we show that Prd maturation requires a small dimeric protein that we have named PrdH. PrdH is co-encoded with the PrdA and PrdB subunits of Prd and also found in species producing similar reductases. By producing stable variants of PrdA and PrdB, we demonstrate that PrdH-mediated cleavage and pyruvoyl formation in the PrdA subunit require PrdB, which can be harnessed to produce active recombinant Prd for subsequent analyses. We further created PrdA- and PrdH-mutants to get insight into the interaction of the components and into the processing reaction itself. Finally, we show that deletion ofprdHinC. difficilerenders the corresponding mutant blind to proline, suggesting that this processing factor is essential for proline utilization. Due to the link between Stickland fermentation and pathogenesis, we suggest PrdH may be an attractive target for drug development.Significance StatementEnergy conservation via Stickland fermentation was first described in the 1930s, yet information about the key enzyme of this process, Prd, is scarce, despite the fact that its central role in both metabolism and toxin production make it a promising potential drug target. Here we show how a small, previously overlooked protein that we named PrdH mediates the formation of the catalytically essential pyruvoyl-group in the active center of Prd.In vivostudies inC. difficileemphasize its critical importance in the utilization of proline. The known interplay between proline reduction and toxin production leads us to suggest PrdH as a potential drug target. Moreover, our findings open the door for further structural and functional studies with recombinantly produced active Prd.

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Protein-Derived Cofactors

Cited by 3 publications

References 158 publications

Plant Copper Metalloenzymes As Prospects for New Metabolism Involving Aromatic Compounds

Plant Copper Metalloenzymes As Prospects for New Metabolism Involving Aromatic Compounds

Formation of the pyruvoyl-dependent proline reductase Prd from Clostridioides difficile requires the maturation enzyme PrdH

Formation of the pyruvoyl-dependent proline reductase Prd fromClostridioides difficilerequires the maturation enzyme PrdH

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