2J8M

Structure of P. aeruginosa acetyltransferase PA4866

2J8M の概要

• エントリーDOI: 10.2210/pdb2j8m/pdb
• 関連するPDBエントリー: 1YVO 2BL1 2J8N 2J8R
• 分子名称: ACETYLTRANSFERASE PA4866 FROM P. AERUGINOSA, AZIDE ION, GLYCEROL, ... (5 entities in total)
• 機能のキーワード: gcn5 family, phosphinothricin, methionine sulfone, transferase methionine sulfoximine, n-acetyl transferase, hypothetical protein, transferase
• 由来する生物種: PSEUDOMONAS AERUGINOSA
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 38448.94

構造登録者

Davies, A.M.,Tata, R.,Beavil, R.L.,Sutton, B.J.,Brown, P.R. (登録日: 2006-10-26, 公開日: 2007-02-06, 最終更新日: 2023-12-13)

主引用文献

Davies, A.M.,Tata, R.,Beavil, R.L.,Sutton, B.J.,Brown, P.R.
L-Methionine Sulfoximine, But not Phosphinothricin, is a Substrate for an Acetyltransferase (Gene Pa4866) from Pseudomonas Aeruginosa: Structural and Functional Studies.
Biochemistry, 46:1829-, 2007

PubMed Abstract: The gene PA4866 from Pseudomonas aeruginosa is documented in the Pseudomonas genome database as encoding a 172 amino acid hypothetical acetyltransferase. We and others have described the 3D structure of this protein (termed pita) [Davies et al. (2005) Proteins: Struct., Funct., Bioinf. 61, 677-679; Nocek et al., unpublished results], and structures have also been reported for homologues from Agrobacterium tumefaciens (Rajashankar et al., unpublished results) and Bacillus subtilis [Badger et al. (2005) Proteins: Struct., Funct., Bioinf. 60, 787-796]. Pita homologues are found in a large number of bacterial genomes, and while the majority of these have been assigned putative phosphinothricin acetyltransferase activity, their true function is unknown. In this paper we report that pita has no activity toward phosphinothricin. Instead, we demonstrate that pita acts as an acetyltransferase using the glutamate analogues l-methionine sulfoximine and l-methionine sulfone as substrates, with Km(app) values of 1.3 +/- 0.21 and 1.3 +/- 0.13 mM and kcat(app) values of 505 +/- 43 and 610 +/- 23 s-1 for l-methionine sulfoximine and l-methionine sulfone, respectively. A high-resolution (1.55 A) crystal structure of pita in complex with one of these substrates (l-methionine sulfoximine) has been solved, revealing the mode of its interaction with the enzyme. Comparison with the apoenzyme structure has also revealed how certain active site residues undergo a conformational change upon substrate binding. To investigate the role of pita in P. aeruginosa, a mutant strain, Depp4, in which pita was inactivated through an in-frame deletion, was constructed by allelic exchange. Growth of strain Depp4 in the absence of glutamine was inhibited by l-methionine sulfoximine, suggesting a role for pita in protecting glutamine synthetase from inhibition.

PubMed: 17253769
DOI: 10.1021/BI0615238

実験手法

X-RAY DIFFRACTION (1.44 Å)