5K45

Wolinella succinogenes L-asparaginase P121 + L-Glutamic acid

5K45 の概要

• エントリーDOI: 10.2210/pdb5k45/pdb
• 関連するPDBエントリー: 5K3O 5K4G 5K4H
• 分子名称: L-asparaginase, GLUTAMIC ACID (3 entities in total)
• 機能のキーワード: l-asparaginase, p121, l-glutamic acid, hydrolase
• 由来する生物種: Wolinella succinogenes (strain ATCC 29543 / DSM 1740 / LMG 7466 / NCTC 11488 / FDC 602W)
• 細胞内の位置: Cytoplasm: P50286
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 140203.48

構造登録者

Nguyen, H.A.,Lave, A. (登録日: 2016-05-20, 公開日: 2017-03-29, 最終更新日: 2023-09-27)

主引用文献

Nguyen, H.A.,Durden, D.L.,Lavie, A.
The differential ability of asparagine and glutamine in promoting the closed/active enzyme conformation rationalizes the Wolinella succinogenes L-asparaginase substrate specificity.
Sci Rep, 7:41643-41643, 2017

PubMed Abstract: Many side effects of current FDA-approved L-asparaginases have been related to their secondary L-glutaminase activity. The Wolinella succinogenes L-asparaginase (WoA) has been reported to be L-glutaminase free, suggesting it would have fewer side effects. Unexpectedly, the WoA variant with a proline at position 121 (WoA-P) was found to have L-glutaminase activity in contrast to Uniprot entry P50286 (WoA-S) that has a serine residue at this position. Towards understanding how this residue impacts the L-glutaminase property, kinetic analysis was coupled with crystal structure determination of these WoA variants. WoA-S was confirmed to have much lower L-glutaminase activity than WoA-P, yet both showed comparable L-asparaginase activity. Structures of the WoA variants in complex with L-aspartic acid versus L-glutamic acid provide insights into their differential substrate selectivity. Structural analysis suggests a mechanism by which residue 121 impacts the conformation of the conserved tyrosine 27, a component of the catalytically-important flexible N-terminal loop. Surprisingly, we could fully model this loop in either its open or closed conformations, revealing the roles of specific residues of an evolutionary conserved motif among this L-asparaginase family. Together, this work showcases critical residues that influence the ability of the flexible N-terminal loop for adopting its active conformation, thereby effecting substrate specificity.

PubMed: 28139703
DOI: 10.1038/srep41643

実験手法

X-RAY DIFFRACTION (1.63 Å)