4GWC

Crystal Structure of Mn2+2,Zn2+-Human Arginase I

4GWC の概要

• エントリーDOI: 10.2210/pdb4gwc/pdb
• 関連するPDBエントリー: 2PHA 4GWD
• 分子名称: Arginase-1, MANGANESE (II) ION, ZINC ION, ... (4 entities in total)
• 機能のキーワード: arginase fold, hydrolase
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Cytoplasm: P05089
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 69910.33

構造登録者

D'Antonio, E.L.,Hai, Y.,Christianson, D.W. (登録日: 2012-09-01, 公開日: 2012-09-26, 最終更新日: 2023-09-13)

主引用文献

D'Antonio, E.L.,Hai, Y.,Christianson, D.W.
Structure and function of non-native metal clusters in human arginase I.
Biochemistry, 51:8399-8409, 2012

PubMed Abstract: Various binuclear metal ion clusters and complexes have been reconstituted in crystalline human arginase I by removing the Mn(2+)(2) cluster of the wild-type enzyme with metal chelators and subsequently soaking the crystalline apoenzyme in buffer solutions containing NiCl(2) or ZnCl(2). X-ray crystal structures of these metal ion variants are correlated with catalytic activity measurements that reveal differences resulting from metal ion substitution. Additionally, treatment of crystalline Mn(2+)(2)-human arginase I with Zn(2+) reveals for the first time the structural basis for inhibition by Zn(2+), which forms a carboxylate-histidine-Zn(2+) triad with H141 and E277. The imidazole side chain of H141 is known to be hyper-reactive, and its chemical modification or mutagenesis is known to similarly compromise catalysis. The reactive substrate analogue 2(S)-amino-6-boronohexanoic acid (ABH) binds as a tetrahedral boronate anion to Mn(2+)(2), Co(2+)(2), Ni(2+)(2), and Zn(2+)(2) clusters in human arginase I, and it can be stabilized by a third inhibitory Zn(2+) ion coordinated by H141. Because ABH binds as an analogue of the tetrahedral intermediate and its flanking transition states in catalysis, this implies that the various metallo-substituted enzymes are capable of some level of catalysis with an actual substrate. Accordingly, we establish the following trend for turnover number (k(cat)) and catalytic efficiency (k(cat)/K(M)): Mn(2+) > Ni(2+) ≈ Co(2+) ≫ Zn(2+). Therefore, Mn(2+) is required for optimal catalysis by human arginase I.

PubMed: 23061982
DOI: 10.1021/bi301145n

実験手法

X-RAY DIFFRACTION (1.9 Å)