4ZU5
Crystal structure of the QdtA 3,4-Ketoisomerase from Thermoanaerobacterium thermosaccharolyticum, apo form
4ZU5 の概要
エントリーDOI | 10.2210/pdb4zu5/pdb |
関連するPDBエントリー | 4ZU4 |
分子名称 | QdtA, THYMIDINE, (2S)-1-[3-{[(2R)-2-hydroxypropyl]oxy}-2,2-bis({[(2R)-2-hydroxypropyl]oxy}methyl)propoxy]propan-2-ol, ... (4 entities in total) |
機能のキーワード | cupin, isomerase, 3, 4-ketoisomerase |
由来する生物種 | Thermoanaerobacterium thermosaccharolyticum |
タンパク質・核酸の鎖数 | 2 |
化学式量合計 | 35184.56 |
構造登録者 | Thoden, J.B.,Vinogradov, E.,Gilbert, M.,Salinger, A.J.,Holden, H.M. (登録日: 2015-05-15, 公開日: 2016-03-30, 最終更新日: 2023-09-27) |
主引用文献 | Thoden, J.B.,Vinogradov, E.,Gilbert, M.,Salinger, A.J.,Holden, H.M. Bacterial Sugar 3,4-Ketoisomerases: Structural Insight into Product Stereochemistry. Biochemistry, 54:4495-4506, 2015 Cited by PubMed Abstract: 3-Acetamido-3,6-dideoxy-d-galactose (Fuc3NAc) and 3-acetamido-3,6-dideoxy-d-glucose (Qui3NAc) are unusual sugars found on the lipopolysaccharides of Gram-negative bacteria and on the S-layers of Gram-positive bacteria. The 3,4-ketoisomerases, referred to as FdtA and QdtA, catalyze the third steps in the respective biosynthetic pathways for these sugars. Whereas both enzymes utilize the same substrate, the stereochemistries of their products are different. Specifically, the hydroxyl groups at the hexose C-4' positions assume the "galactose" and "glucose" configurations in the FdtA and QdtA products, respectively. In 2007 we reported the structure of the apoform of FdtA from Aneurinibacillus thermoaerophilus, which was followed in 2014 by the X-ray analysis of QdtA from Thermoanaerobacterium thermosaccharolyticum as a binary complex. Both of these enzymes belong to the cupin superfamily. Here we report a combined structural and enzymological study to explore the manner in which these enzymes control the stereochemistry of their products. Various site-directed mutant proteins of each enzyme were constructed, and their dTDP-sugar products were analyzed by NMR spectroscopy. In addition, the kinetic parameters for these protein variants were measured, and the structure of one, namely, the QdtA Y17R/R97H double mutant form, was determined to 2.3-Å resolution. Finally, in an attempt to obtain a model of FdtA with a bound dTDP-linked sugar, the 3,4-ketoisomerase domain of a bifunctional enzyme from Shewanella denitrificans was cloned, purified, and crystallized in the presence of a dTDP-linked sugar analogue. Taken together, the results from this investigation demonstrate that it is possible to convert a "galacto" enzyme into a "gluco" enzyme and vice versa. PubMed: 26125548DOI: 10.1021/acs.biochem.5b00541 主引用文献が同じPDBエントリー |
実験手法 | X-RAY DIFFRACTION (1.8 Å) |
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