4BPM

Crystal structure of a human integral membrane enzyme

4BPM の概要

• エントリーDOI: 10.2210/pdb4bpm/pdb
• 関連するPDBエントリー: 4BPD
• 分子名称: PROSTAGLANDIN E SYNTHASE, FUSION PEPTIDE, GLUTATHIONE, 2-[[2,6-bis(chloranyl)-3-[(2,2-dimethylpropanoylamino)methyl]phenyl]amino]-1-methyl-6-(2-methyl-2-oxidanyl-propoxy)-N-[2,2,2-tris(fluoranyl)ethyl]benzimidazole-5-carboxamide, ... (4 entities in total)
• 機能のキーワード: isomerase, cancer, drug target, in meso crystallization, inflammation, inhibitor, leukotriene c4 synthase, lipid metabolism, membrane-associated proteins in eicosanoid and glutathione metabolism, mapag, membrane protein, mpges1, pain, microcrystal, anomalous dispersion, sulfur-sad, s-sad
• 由来する生物種: HOMO SAPIENS; 詳細
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 21286.73

構造登録者

Li, D.,Wang, M.,Olieric, V.,Caffrey, M. (登録日: 2013-05-27, 公開日: 2014-04-16, 最終更新日: 2024-05-08)

主引用文献

Li, D.,Howe, N.,Dukkipati, A.,Shah, S.T.A.,Bax, B.D.,Edge, C.,Bridges, A.,Hardwicke, P.,Singh, O.M.P.,Giblin, G.,Pautsch, A.,Pfau, R.,Schnapp, G.,Wang, M.,Olieric, V.,Caffrey, M.
Crystallizing Membrane Proteins in the Lipidic Mesophase. Experience with Human Prostaglandin E2 Synthase 1 and an Evolving Strategy.
Cryst.Growth Des., 14:2034-, 2014

PubMed Abstract: The lipidic mesophase or in meso method for crystallizing membrane proteins has several high profile targets to its credit and is growing in popularity. Despite its success, the method is in its infancy as far as rational crystallogenesis is concerned. Consequently, significant time, effort, and resources are still required to generate structure-grade crystals, especially with a new target type. Therefore, a need exists for crystallogenesis protocols that are effective with a broad range of membrane protein types. Recently, a strategy for crystallizing a prokaryotic α-helical membrane protein, diacylglycerol kinase (DgkA), by the in meso method was reported (Cryst. Growth. Des.2013, 14, 2846-2857). Here, we describe its application to the human α-helical microsomal prostaglandin E2 synthase 1 (mPGES1). While the DgkA strategy proved useful, significant modifications were needed to generate structure-quality crystals of this important therapeutic target. These included protein engineering, using an additive phospholipid in the hosting mesophase, performing multiple rounds of salt screening, and carrying out trials at 4 °C in the presence of a tight binding ligand. The crystallization strategy detailed here should prove useful for generating structures of other integral membrane proteins by the in meso method.

PubMed: 24803849
DOI: 10.1021/CG500157X

実験手法

X-RAY DIFFRACTION (2.08 Å)