6XRC

Apo NIS synthetase DesD variant R306Q

6XRC の概要

• エントリーDOI: 10.2210/pdb6xrc/pdb
• 関連するPDBエントリー: 6NL2
• 分子名称: Desferrioxamine E biosynthesis protein DesD, GLYCEROL, ADENOSINE-5'-TRIPHOSPHATE, ... (5 entities in total)
• 機能のキーワード: nis synthetase, catalytic variant, siderophore synthesis, biosynthetic protein
• 由来する生物種: Streptomyces coelicolor
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 136776.20

構造登録者

Hoffmann, K.M. (登録日: 2020-07-11, 公開日: 2020-10-28, 最終更新日: 2023-10-18)

主引用文献

Hoffmann, K.M.,Goncuian, E.S.,Karimi, K.L.,Amendola, C.R.,Mojab, Y.,Wood, K.M.,Prussia, G.A.,Nix, J.,Yamamoto, M.,Lathan, K.,Orion, I.W.
Cofactor Complexes of DesD, a Model Enzyme in the Virulence-related NIS Synthetase Family.
Biochemistry, 59:3427-3437, 2020

PubMed Abstract: The understudied nonribosomal-peptide-synthetase-independent siderophore (NIS) synthetase family has been increasingly associated with virulence in bacterial species due to its key role in the synthesis of hydroxamate and carboxylate "stealth" siderophores. We have identified a model family member, DesD, from , to structurally characterize using a combination of a wild-type and a Arg306Gln variant in , cofactor product AMP-bound, and cofactor reactant ATP-bound complexes. The kinetics in the family has been limited by solubility and reporter assays, so we have developed a label-free kinetics assay utilizing a single-injection isothermal-titration-calorimetry-based method. We report second-order rate constants that are 50 times higher than the previous estimations for DesD. Our Arg306Gln DesD variant was also tested under identical buffer and substrate conditions, and its undetectable activity was confirmed. These are the first reported structures for DesD, and they describe the critical cofactor coordination. This is also the first label-free assay to unambiguously determine the kinetics for an NIS synthetase.

PubMed: 32885650
DOI: 10.1021/acs.biochem.9b00899

実験手法

X-RAY DIFFRACTION (2.45 Å)