Reaction Mechanism of Prephenate Dehydrogenase from the Alternative Tyrosine Biosynthesis Pathway in Plants
| Autor: | Joseph M. Jez, Cynthia K. Holland |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification biology Organic Chemistry food and beverages Active site Prephenate dehydrogenase Biochemistry Cofactor Amino acid Enzyme catalysis 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology chemistry Biosynthesis Aromatic amino acids biology.protein Molecular Medicine Tyrosine Molecular Biology |
| Zdroj: | ChemBioChem. 19:1132-1136 |
| ISSN: | 1439-7633 1439-4227 |
| Popis: | Unlike metazoans, plants, bacteria, and fungi retain the enzymatic machinery necessary to synthesize the three aromatic amino acids l-phenylalanine, l-tyrosine, and l-tryptophan de novo. In legumes, such as soybean, alfalfa, and common bean, prephenate dehydrogenase (PDH) catalyzes the tyrosine-insensitive biosynthesis of 4-hydroxyphenylpyruvate, a precursor to tyrosine. The three-dimensional structure of soybean PDH1 was recently solved in complex with the NADP+ cofactor. This structure allowed for the identification of both the cofactor- and ligand-binding sites. Here, we present steady-state kinetic analysis of twenty site-directed active-site mutants of soybean (Glycine max) PDH compared to wild-type. Molecular docking of the substrate, prephenate, into the active site of the enzyme revealed its potential interactions with the active site residues and made a case for the importance of each residue in substrate recognition and/or catalysis, most likely through transition state stabilization. Overall, these results suggested that the active site of the enzyme is highly sensitive to any changes, as even subtle alterations substantially reduced the catalytic efficiency of the enzyme. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text