7VID

The crystal structure of L-leucine dehydrogenase from Pseudomonas aeruginosa

7VID の概要

• エントリーDOI: 10.2210/pdb7vid/pdb
• 分子名称: Leucine dehydrogenase, GLYCEROL (3 entities in total)
• 機能のキーワード: pseudomonas aeruginosa, pa3418, leucine dehydrogenase, branched chain amino acid, oxidoreductase
• 由来する生物種: Pseudomonas aeruginosa PAO1
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 73184.79

構造登録者

Kim, S.,Kang, W.,Yang, J.K. (登録日: 2021-09-26, 公開日: 2022-06-29, 最終更新日: 2023-11-29)

主引用文献

Kim, S.,Koh, S.,Kang, W.,Yang, J.K.
The Crystal Structure of L-Leucine Dehydrogenase from Pseudomonas aeruginosa.
Mol.Cells, 45:495-501, 2022

PubMed Abstract: Leucine dehydrogenase (LDH, EC 1.4.1.9) catalyzes the reversible deamination of branched-chain L-amino acids to their corresponding keto acids using NAD as a cofactor. LDH generally adopts an octameric structure with symmetry, generating a molecular mass of approximately 400 kDa. Here, the crystal structure of the LDH from (-LDH) was determined at 2.5 Å resolution. Interestingly, the crystal structure shows that the enzyme exists as a dimer with symmetry in a crystal lattice. The dimeric structure was also observed in solution using multiangle light scattering coupled with size-exclusion chromatography. The enzyme assay revealed that the specific activity was maximal at 60°C and pH 8.5. The kinetic parameters for three different amino acid and the cofactor (NAD) were determined. The crystal structure represents that the subunit has more compact structure than homologs' structure. In addition, the crystal structure along with sequence alignments indicates a set of non-conserved arginine residues which are important in stability. Subsequent mutation analysis for those residues revealed that the enzyme activity reduced to one third of the wild type. These results provide structural and biochemical insights for its future studies on its application for industrial purposes.

PubMed: 35698914
DOI: 10.14348/molcells.2022.0012

実験手法

X-RAY DIFFRACTION (2.5 Å)