4ECA

ASPARAGINASE FROM E. COLI, MUTANT T89V WITH COVALENTLY BOUND ASPARTATE

4ECA の概要

• エントリーDOI: 10.2210/pdb4eca/pdb
• 分子名称: L-ASPARAGINE AMIDOHYDROLASE (2 entities in total)
• 機能のキーワード: hydrolase, acyl-enzyme intermediate, threonine amidohydrolase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 138959.70

構造登録者

Palm, G.J.,Lubkowski, J.,Wlodawer, A. (登録日: 1997-02-21, 公開日: 1997-06-16, 最終更新日: 2023-08-09)

主引用文献

Palm, G.J.,Lubkowski, J.,Derst, C.,Schleper, S.,Rohm, K.H.,Wlodawer, A.
A covalently bound catalytic intermediate in Escherichia coli asparaginase: crystal structure of a Thr-89-Val mutant.
FEBS Lett., 390:211-216, 1996

PubMed Abstract: Escherichia coli asparaginase II catalyzes the hydrolysis of L-asparagine to L-aspartate via a threonine-bound acyl-enzyme intermediate. A nearly inactive mutant in which one of the active site threonines, Thr-89, was replaced by valine was constructed, expressed, and crystallized. Its structure, solved at 2.2 A resolution, shows high overall similarity to the wild-type enzyme, but an aspartyl moiety is covalently bound to Thr-12, resembling a reaction intermediate. Kinetic analysis confirms the deacylation deficiency, which is also explained on a structural basis. The previously identified oxyanion hole is described in more detail.

PubMed: 8706862
DOI: 10.1016/0014-5793(96)00660-6

実験手法

X-RAY DIFFRACTION (2.2 Å)