3TKJ

Crystal Structure of Human Asparaginase-like Protein 1 Thr168Ala

3TKJ の概要

• エントリーDOI: 10.2210/pdb3tkj/pdb
• 関連するPDBエントリー: 3TKX
• 分子名称: L-asparaginase, SODIUM ION, TRIS(HYDROXYETHYL)AMINOMETHANE, ... (5 entities in total)
• 機能のキーワード: alpha-beta-beta-alpha, hydrolase
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Cytoplasm: Q7L266
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 66921.75

構造登録者

Li, W.Z.,Yogesha, S.D.,Liu, J.,Zhang, Y. (登録日: 2011-08-26, 公開日: 2012-10-10, 最終更新日: 2024-02-28)

主引用文献

Li, W.,Cantor, J.R.,Yogesha, S.D.,Yang, S.,Chantranupong, L.,Liu, J.Q.,Agnello, G.,Georgiou, G.,Stone, E.M.,Zhang, Y.
Uncoupling Intramolecular Processing and Substrate Hydrolysis in the N-Terminal Nucleophile Hydrolase hASRGL1 by Circular Permutation.
Acs Chem.Biol., 7:1840-1847, 2012

PubMed Abstract: The human asparaginase-like protein 1 (hASRGL1) catalyzes the hydrolysis of l-asparagine and isoaspartyl-dipeptides. As an N-terminal nucleophile (Ntn) hydrolase superfamily member, the active form of hASRGL1 is generated by an intramolecular cleavage step with Thr168 as the catalytic residue. However, in vitro, autoprocessing is incomplete (~50%), fettering the biophysical characterization of hASRGL1. We circumvented this obstacle by constructing a circularly permuted hASRGL1 that uncoupled the autoprocessing reaction, allowing us to kinetically and structurally characterize this enzyme and the precursor-like hASRGL1-Thr168Ala variant. Crystallographic and biochemical evidence suggest an activation mechanism where a torsional restraint on the Thr168 side chain helps drive the intramolecular processing reaction. Cleavage and formation of the active site releases the torsional restriction on Thr168, which is facilitated by a small conserved Gly-rich loop near the active site that allows the conformational changes necessary for activation.

PubMed: 22891768
DOI: 10.1021/cb300232n

実験手法

X-RAY DIFFRACTION (2.3 Å)