Expression, purification, crystallization and preliminary X-ray analysis of perakine reductase, a new member of the aldo-keto reductase enzyme superfamily from higher plants

Rosenthal, Cindy; Muêller, U.; Panjikar, Santosh; Sun, Lianli; Ruppert, Martin; Zhao, Yu; Stöckigt, Joachim

doi:10.1107/s174430910605041x

Cited by 15 publications

(11 citation statements)

References 24 publications

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“…Several refined [STR (Ma et al, 2006) and SGD ] and partial [raucaffricine glucosidase , perakine reductase (Rosenthal et al, 2006) and vinorine synthase ] crystal structures for enzymes involved in MIA biosynthesis in R. serpentina have recently been reported (Figure 2). The information obtained from the crystal structure of STR complexed with strictosidine was used to produce the rational site-directed V208A mutation of STR.…”

Section: Metabolic Engineering Of Substrate Specificity Of Mia Pathwamentioning

confidence: 99%

Opium poppy and Madagascar periwinkle: model non‐model systems to investigate alkaloid biosynthesis in plants

Facchini¹,

Luca²

2008

The Plant Journal

224

134

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SummaryAlkaloids represent a large and diverse group of compounds that are related by the occurrence of a nitrogen atom within a heterocyclic backbone. Unlike other types of secondary metabolites, the various structural categories of alkaloids are unrelated in terms of biosynthesis and evolution. Although the biology of each group is unique, common patterns have become apparent. Opium poppy (Papaver somniferum), which produces several benzylisoquinoline alkaloids, and Madagascar periwinkle (Catharanthus roseus), which accumulates an array of monoterpenoid indole alkaloids, have emerged as the premier organisms used to study plant alkaloid metabolism. The status of these species as model systems results from decades of research on the chemistry, enzymology and molecular biology responsible for the biosynthesis of valuable pharmaceutical alkaloids. Opium poppy remains the only commercial source for morphine, codeine and semisynthetic analgesics, such as oxycodone, derived from thebaine. Catharanthus roseus is the only source for the anti-cancer drugs vinblastine and vincristine. Impressive collections of cDNAs encoding biosynthetic enzymes and regulatory proteins involved in the formation of benzylisoquinoline and monoterpenoid indole alkaloids are now available, and the rate of gene discovery has accelerated with the application of genomics. Such tools have allowed the establishment of models that describe the complex cell biology of alkaloid metabolism in these important medicinal plants. A suite of biotechnological resources, including genetic transformation protocols, has allowed the application of metabolic engineering to modify the alkaloid content of these and related species. An overview of recent progress on benzylisoquinoline and monoterpenoid indole alkaloid biosynthesis in opium poppy and C. roseus is presented.

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Section: Metabolic Engineering Of Substrate Specificity Of Mia Pathwamentioning

confidence: 99%

Opium poppy and Madagascar periwinkle: model non‐model systems to investigate alkaloid biosynthesis in plants

Facchini¹,

Luca²

2008

The Plant Journal

224

134

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“…The absence of both a Rossmann fold (11) and the catalytic motif Tyr-Xaa-Xaa-Xaa-Lys (12) (13). Furthermore, sequence alignment and preliminary x-ray analysis of a methylated PR mutant support the view that PR represents a novel member of the AKR superfamily (6,14). In fact, PR would be the first example of a plant AKR involved in indole alkaloid biosynthesis in contrast to the plant AKR-type codeinone reductase, which is involved in isoquinoline alkaloid metabolism (15).…”

mentioning

confidence: 77%

“…Bacteria were grown at 25°C for large scale enzyme production, and the conditions for cultivation, induction and purification of recombinant PR were the same as previously reported (6,14).…”

Section: Methodsmentioning

confidence: 99%

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Crystal Structure of Perakine Reductase, Founding Member of a Novel Aldo-Keto Reductase (AKR) Subfamily That Undergoes Unique Conformational Changes during NADPH Binding

Sun

Chen

Rajendran

et al. 2012

Journal of Biological Chemistry

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Background: Perakine reductase (PR) is an AKR involved in the Rauvolfia alkaloid biosynthetic network. Results: Three-dimensional structures of PR and the A213W mutant complex with NADPH were solved. Conclusion: PR folds as an unusual ␣ 8 /␤ 6 barrel and undergoes unexpected conformational changes upon NADPH binding. Significance: PR represents the founding member of the new AKR13D subfamily and provides a structural and cofactor binding template for the AKR13 family.

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“…The availability of crystal structures for various enzymes involved in MIA biosynthesis, namely STR, SGD, raucaffricine glucosidase, perakine reductase and vinorine synthase (Ma et al, , 2006Rosenthal et al, 2006;Ruppert et al, 2006;Barleben et al, 2007), have set the stage for rational protein engineering and for altering the substrate specificity of these reactions. In the case of STR, crystal structures were produced with the enzyme complexed to secologanin or to tryptamine.…”

Section: Metabolic Engineering Using Enzymes With Altered Substrate Smentioning

confidence: 99%

Monoterpenoid Indole Alkaloid Biosynthesis

Luca¹

2011

Plant Metabolism and Biotechnology

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A unique feature of MIA assembly is the participation of separate pathways that distinguish tryptamine from the tryptophan branch of the shikimate pathway and secologanin from the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway rather than the mevalonic acid (MVA) pathway (Figure 10.2). Many biochemical studies on tryptophan decarboxylase (TDC) involved in tryptamine formation from tryptophan have been conducted with Catharanthus roseus cell cultures and in intact plants. The cDNA for this gene was the first to be identified and functionally characterised in Catharanthus , with the recombinant protein being able to decarboxylate tryptophan but not tyrosine or phenylalanine. Labelling studies using Catharanthus roseus (Contin et al., 1998) and Ophiorrhiza pumila (Yamazaki et al., 2004) cell cultures with [1-13 C]-glucose showed that the MEP pathway, rather than the mevalonic pathway, contributes to the formation of this key precursor. While many of the Catharanthus genes for the MEP pathway, including deoxyxylulose 5-phosphate synthase (DXS), deoxyxylulose 5-phosphate reductoisomerase (DXR), CO 2 Primary Carbon Metabolism 3PGA + Pyruvate Pyruvate E-4-P + PEP Plastid Shikimate Pathway Plastid MEP Pathway Cytosol Mevalonic acid Pathway ? Sesquiterpenes, Triterpenes MIAs Monoterpenes Diterpenes Tetraterpenes Figure 10.2 Linking primary metabolism and MIA biosynthesis. Primary metabolism converts CO 2 fixed during photosynthesis to erythrose-4-phosphate (E-4-P), phospoenolpyruvate (PEP), 3-phosphoglycerate (3PGA) and pyruvate which are used to elaborate the plastid-localised shikimate and MEP pathways together with the cytosol localised mevalonic acid pathway for biosynthesis of MIAs, as well as different terpenes. The shikimate pathway provides the tryptophan that is then converted in the cytosol to tryptamine which is utilised in the biosynthesis of MIAs. It is not clear how much crosstalk takes place between the MEP and mevalonic acid pathways

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Expression, purification, crystallization and preliminary X-ray analysis of perakine reductase, a new member of the aldo-keto reductase enzyme superfamily from higher plants

Cited by 15 publications

References 24 publications

Opium poppy and Madagascar periwinkle: model non‐model systems to investigate alkaloid biosynthesis in plants

Opium poppy and Madagascar periwinkle: model non‐model systems to investigate alkaloid biosynthesis in plants

Crystal Structure of Perakine Reductase, Founding Member of a Novel Aldo-Keto Reductase (AKR) Subfamily That Undergoes Unique Conformational Changes during NADPH Binding

Monoterpenoid Indole Alkaloid Biosynthesis

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