Arogenate dehydratase (ADT) (EC 4.2.1.91) is an enzyme that catalyzes the chemical reaction. Hence, this enzyme has one substrate, L-arogenate, but 3 products:L-phenylalanine, H2O, and CO2. Certain forms of the protein have the potential to catalyze a second reaction. This enzyme participates in phenylalanine, tyrosine, and tryptophan biosynthesis. The carboxyl and hydroxide groups attached to the 2,5-cyclohexene ring are eliminated from L-arogenate, leaving as carbon dioxide and water. The 2,5-cyclohexene ring becomes a phenyl ring, and L-phenylalanine is formed. Certain forms of ADT have been shown to exhibit some prephenate dehydratase (PDT) activity in addition to the standard ADT activity described above. Known as cyclohexadienyl dehydratases or carbocyclohexadienyl dehydratases, these forms of the enzyme catalyze the same type of reaction (a decarboxylation and a dehydration) on prephenate. The carboxyl and hydroxide groups attached to the 2,5-cyclohexene ring are removed, leaving phenylpyruvate. ADT catalyzes a reaction categorized by two major changes in the structure of the substrate, these being a decarboxylation and a dehydration; the enzyme removes a carboxyl group and a water molecule (respectively). Both potential products of this reaction (L-arogenate and phenylpyruvate) occur at or near the end of the biosynthetic pathway. Total synthesis of L-arogenate has been reported.
Background References
1. Cho, Man-Ho; Corea, Oliver R. A.; Yang, Hong; Bedgar, Diana L.; Laskar, Dhrubojyoti D.; Anterola, Aldwin M.; et al. (2007). "Phenylalanine biosynthesis in Arabidopsis thaliana-- identification and characterization of arogenate dehydratases". J. Biol. Chem. 282 (42): 30827–35.
2. Maeda, H.; Shasany, A. K.; Schnepp, J.; Orlova, I.; Taguchi, G.; Cooper, B. R.; et al. (2010). "RNAi suppression of Arogenate Dehydratase 1 reveals that phenylalanine is synthesized predominantly via the arogenate pathway in petunia petals". Plant Cell. 22 (3): 832–849.
Western blot analysis of Arogenate dehydratase on rice tissue lysate with Rabbit anti-Arogenate dehydratase antibody (ER1901-51) at 1/100 dilution.
Lysates/proteins at 15 µg/Lane. Exposure time: 1 minutes; ECL: K1801
Blocking: 5% BSA in PBS, 1 hour at room temperature Primary antibody: ER1901-51, 1/100 in 5% BSA at room temperature for 2 hours Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/5000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 40 kDa Observed band size: 40 kDa
Immunohistochemical analysis of paraffin-embedded rice tissue using anti-Arogenate dehydratase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-51, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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