1CZE

ASPARTATE AMINOTRANSFERASE MUTANT ATB17/139S/142N WITH SUCCINIC ACID

1CZE の概要

• エントリーDOI: 10.2210/pdb1cze/pdb
• 関連するPDBエントリー: 1YOO
• 分子名称: ASPARTATE AMINOTRANSFERASE, PYRIDOXAL-5'-PHOSPHATE, SUCCINIC ACID, ... (4 entities in total)
• 機能のキーワード: aspartate aminotransferase, substrate specificity, transferase
• 由来する生物種: Escherichia coli
• 細胞内の位置: Cytoplasm: P00509
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 44027.44

構造登録者

Okamoto, A.,Oue, S.,Yano, T.,Kagamiyama, H. (登録日: 1999-09-02, 公開日: 2000-02-28, 最終更新日: 2023-08-09)

主引用文献

Oue, S.,Okamoto, A.,Yano, T.,Kagamiyama, H.
Cocrystallization of a mutant aspartate aminotransferase with a C5-dicarboxylic substrate analog: structural comparison with the enzyme-C4-dicarboxylic analog complex.
J.Biochem.(Tokyo), 127:337-343, 2000

PubMed Abstract: A mutant Escherichia coil aspartate aminotransferase with 17 amino acid substitutions (ATB17), previously created by directed evolution, shows increased activity for beta-branched amino acids and decreased activity for the native substrates, aspartate and glutamate. A new mutant (ATBSN) was generated by changing two of the 17 mutated residues back to the original ones. ATBSN recovered the activities for aspartate and glutamate to the level of the wild-type enzyme while maintaining the enhanced activity of ATB17 for the other amino acid substrates. The absorption spectrum of the bound coenzyme, pyridoxal 5'-phosphate, also returned to the original state. ATBSN shows significantly increased affinity for substrate analogs including succinate and glutarate, analogs of aspartate and glutamate, respectively. Hence, we could cocrystallize ATBSN with succinate or glutarate, and the structures show how the enzyme can bind two kinds of dicarboxylic substrates with different chain lengths. The present results may also provide an insight into the long-standing controversies regarding the mode of binding of glutamate to the wild-type enzyme.

PubMed: 10731702

実験手法

X-RAY DIFFRACTION (2.4 Å)