Modulation of flavonoid metabolism in Arabidopsis using a phage-derived antibody

Santos, Michael O.; Crosby, William L.; Winkel, Brenda S.J.

doi:10.1023/b:molb.0000034088.64105.13

Cited by 10 publications

(7 citation statements)

References 43 publications

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“…Single‐chain Fv antibodies selected by phage display can be used to block enzyme activity in animals and plants (Lener et al ., 2000; Jobling et al ., 2003; Santos et al ., 2004). We used this approach to generate high‐affinity scFvs specific to human ODC to immunomodulate polyamine biosynthesis in human cells.…”

Section: Resultsmentioning

confidence: 99%

“…We have demonstrated that immunomodulation is a powerful tool for the analysis of biochemical pathways. Until now, there have only been two successful reports of antibody‐based metabolic engineering, involving the inhibition of potato starch branching enzyme A (SBE A) activity and the modulation of flavonoid metabolism in Arabidopsis (Jobling et al ., 2003; Santos et al ., 2004). In the present study, constitutive expression of an ODC‐specific scFv in the plant cytosol led to a reduction in ODC activity in vivo , decreased levels of putrescine, spermidine and spermine and an aberrant morphological phenotype.…”

Section: Resultsmentioning

confidence: 99%

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Immunomodulation of polyamine biosynthesis in tobacco plants has a significant impact on polyamine levels and generates a dwarf phenotype

Nölke

Schneider

Fischer

et al. 2005

Plant Biotechnology Journal

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SummaryOrnithine decarboxylase (ODC) is a cytosolic enzyme that catalyses the direct decarboxylation of L -ornithine to putrescine, one of the rate-limiting steps of polyamine biosynthesis in plants. In the present study, an ODC-specific murine single-chain antibody fragment (scFvODC1) was generated by phage display technology. To evaluate the effect of the recombinant antibody fragment on ODC activity and polyamine levels, we produced transgenic tobacco plants that accumulated scFvODC1 in the cytosol. Expression levels of up to 4% total soluble protein (TSP) were achieved, resulting in the inhibition of up to 90% of endogenous ODC activity. A significant reduction in putrescine, spermidine and spermine levels was observed in transgenic lines producing high levels of scFvODC1. Furthermore, these lines showed developmental abnormalities and a dwarf phenotype. We show that the immunomodulation of enzyme activity is a powerful approach that can be used to alter complex and tightly controlled metabolic pathways, allowing specific steps in the pathway to be blocked and the resulting physiological effects to be investigated.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Immunomodulation of polyamine biosynthesis in tobacco plants has a significant impact on polyamine levels and generates a dwarf phenotype

Nölke

Schneider

Fischer

et al. 2005

Plant Biotechnology Journal

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“…Thus, camelid single-domain antibodies have been demonstrated to be stable even at reducing conditions and could inhibit enzymes quiet efficiently. Other successful experiments describe the modulation of the flavonoid metabolism and the immunomodulation of the polyamine biosynthesis using scFv intrabodies inhibiting enzymes (28,29) . ScFvs against small heat shock proteins (sHSP) have been expressed in the cytosol of transgenic tobacco plants to prevent the assembly of heat stress granula.…”

Section: Techniques and Constructs To Efficiently Transform Plants Anmentioning

confidence: 99%

Immunomodulation of Plant Function by In Vitro Selected Single-Chain Fv Intrabodies

Gahrtz

Conrad

2009

Recombinant Proteins From Plants

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In this chapter, we discuss and compare the different concepts and examples as well as present the basic protocols for applying intrabody-based approaches in plants for the investigation of cell functions and plant cell-pathogen interactions. The immunomodulation strategy, a molecular technique that allows to interfere with cellular metabolism, signal transduction pathways, or pathogen infectivity, is based on the ectopic expression of genes encoding specific recombinant antibodies. This needs basic prerequisites to be successfully applied as resources and techniques to isolate specific recombinant antibodies with sufficient binding parameters to bind and to block even low-concentrated targets or to compete successfully with substrates and ligands. Also techniques and constructs to efficiently transform plants and to target recombinant antibodies to selected compartments are important requirements. Basic protocols for all these techniques are provided.

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“…In most cytosolic expressions of scFvs. levels were found to be very low or just at the detection limit (Xiao et cd., 2000;Santos et al, 2004). A notable exception is an scFv antibody isolated by phage display that accumulated up to 1.0% of TSP in the plant cytosol (de Jaeger et al, 1998).…”

Section: Stable Expression In Plantsmentioning

confidence: 99%

Plantibodies: A Novel Strategy to Create Pathogen-Resistant Plants

Liao

Zhao

et al. 2006

Biotechnology and Genetic Engineering Reviews

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Modulation of flavonoid metabolism in Arabidopsis using a phage-derived antibody

Cited by 10 publications

References 43 publications

Immunomodulation of polyamine biosynthesis in tobacco plants has a significant impact on polyamine levels and generates a dwarf phenotype

Immunomodulation of polyamine biosynthesis in tobacco plants has a significant impact on polyamine levels and generates a dwarf phenotype

Immunomodulation of Plant Function by In Vitro Selected Single-Chain Fv Intrabodies

Plantibodies: A Novel Strategy to Create Pathogen-Resistant Plants

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