5G3P

Bacillus cereus formamidase (BceAmiF) acetylated at the active site.

5G3P の概要

• エントリーDOI: 10.2210/pdb5g3p/pdb
• 関連するPDBエントリー: 5G3O
• 分子名称: FORMAMIDASE, DI(HYDROXYETHYL)ETHER, 1,2-ETHANEDIOL, ... (9 entities in total)
• 機能のキーワード: hydrolase, formamidase, amidase, nitrilase superfamily
• 由来する生物種: BACILLUS CEREUS
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 235620.45

構造登録者

Gavira, J.A.,Conejero-Muriel, M.,Martinez-Rodriguez, S. (登録日: 2016-04-29, 公開日: 2017-04-12, 最終更新日: 2024-11-13)

主引用文献

Martinez-Rodriguez, S.,Conejero-Muriel, M.,Gavira, J.A.
A novel cysteine carbamoyl-switch is responsible for the inhibition of formamidase, a nitrilase superfamily member.
Arch.Biochem.Biophys., 662:151-159, 2019

PubMed Abstract: Formamidases (EC 3.5.1.49) and amidases (EC 3.5.1.4) are paralogous cysteine-dependent enzymes which catalyze the conversion of amide substrates to ammonia and the corresponding carboxylic acid. Both enzymes have been suggested as an alternative pathway for ammonia production during urea shortage. Urea was proved key in the transcriptional regulation of formamidases/amidases, connecting urea level to amide metabolism. In addition, different amidases have also been shown to be inhibited by urea, pointing to urea-regulation at the enzymatic level. Although amidases have been widely studied due to its biotechnological application in the hydrolysis of aliphatic amides, up to date, only two formamidases have been extensively characterized, belonging to Helicobacter pylori (HpyAmiF) and Bacillus cereus (BceAmiF). In this work, we report the first structure of an acyl-intermediate of BceAmiF. We also report the inhibition of BceAmiF by urea, together with mass spectrometry studies confirming the S-carbamoylation of BceAmiF after urea treatment. X-ray studies of urea-soaked BceAmiF crystals showed short- and long-range rearrangements affecting oligomerization interfaces. Since cysteine-based switches are known to occur in the regulation of different metabolic and signaling pathways, our results suggest a novel S-carbamoylation-switch for the regulation of BceAmiF. This finding could relate to previous observations of unexplained modifications in the catalytic cysteine of different nitrilase superfamily members and therefore extending this regulation mechanism to the whole nitrilase superfamily.

PubMed: 30528776
DOI: 10.1016/j.abb.2018.12.008

実験手法

X-RAY DIFFRACTION (1.78 Å)