Opposite stereoselectivities of dirigent proteins in Arabidopsis and schizandra species

Kye-Won Kim; Syed G A Moinuddin; Kathleen M Atwell; Michael A Costa; Laurence B Davin; Norman G Lewis

doi:10.1074/jbc.M112.387423

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Opposite stereoselectivities of dirigent proteins in Arabidopsis and schizandra species

Journal article

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Opposite stereoselectivities of dirigent proteins in Arabidopsis and schizandra species

Kye-Won Kim, Syed G A Moinuddin, Kathleen M Atwell, Michael A Costa, Laurence B Davin and Norman G Lewis

The Journal of biological chemistry, Vol.287(41), pp.33957-33972

10/05/2012

DOI: https://doi.org/10.1074/jbc.M112.387423

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https://hdl.handle.net/2376/105780

PMCID: PMC3464507

PMID: 22854967

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Abstract

Furans - metabolism

Arabidopsis - chemistry

Plant Roots - metabolism

Schisandra - metabolism

Species Specificity

Schisandra - chemistry

Plant Roots - genetics

Substrate Specificity

Lignans - metabolism

Arabidopsis - metabolism

Arabidopsis - genetics

Sequence Homology, Amino Acid

Plant Proteins - genetics

Plant Proteins - chemistry

Plant Roots - chemistry

Plant Proteins - metabolism

Binding Sites

Schisandra - genetics

How stereoselective monolignol-derived phenoxy radical-radical coupling reactions are differentially biochemically orchestrated in planta, whereby for example they afford (+)- and (-)-pinoresinols, respectively, is both a fascinating mechanistic and evolutionary question. In earlier work, biochemical control of (+)-pinoresinol formation had been established to be engendered by a (+)-pinoresinol-forming dirigent protein in Forsythia intermedia, whereas the presence of a (-)-pinoresinol-forming dirigent protein was indirectly deduced based on the enantiospecificity of downstream pinoresinol reductases (AtPrRs) in Arabidopsis thaliana root tissue. In this study of 16 putative dirigent protein homologs in Arabidopsis, AtDIR6, AtDIR10, and AtDIR13 were established to be root-specific using a β-glucuronidase reporter gene strategy. Of these three, in vitro analyses established that only recombinant AtDIR6 was a (-)-pinoresinol-forming dirigent protein, whose physiological role was further confirmed using overexpression and RNAi strategies in vivo. Interestingly, its closest homolog, AtDIR5, was also established to be a (-)-pinoresinol-forming dirigent protein based on in vitro biochemical analyses. Both of these were compared in terms of properties with a (+)-pinoresinol-forming dirigent protein from Schizandra chinensis. In this context, sequence analyses, site-directed mutagenesis, and region swapping resulted in identification of putative substrate binding sites/regions and candidate residues controlling distinct stereoselectivities of coupling modes.

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Title: Opposite stereoselectivities of dirigent proteins in Arabidopsis and schizandra species
Creators: Kye-Won Kim - Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA
Syed G A Moinuddin
Kathleen M Atwell
Michael A Costa
Laurence B Davin
Norman G Lewis
Publication Details: The Journal of biological chemistry, Vol.287(41), pp.33957-33972
Academic Unit: Biological Chemistry, Institute of
Publisher: United States
Grant note: T32 GM008336 / NIGMS NIH HHS
Identifiers: 99900546702701842
Language: English
Resource Type: Journal article