tailieunhanh - Báo cáo khoa học: Sensor of phospholipids inStreptomycesphospholipase D

Recently, we identified Ala426 and Lys438 of phospholipase D fromStrep-tomyces septatus TH-2 (TH-2PLD) as important residues for activity, sta-bility and selectivity in transphosphatidylation. These residues are located in a C-terminal flexible loop separate from two catalytic HxKxxxxD motifs. | ễFEBS Journal Sensor of phospholipids in Streptomyces phospholipase D Yoshiko Uesugi Jiro Arima Masaki Iwabuchi and Tadashi Hatanaka Research Institute for BiologicalSciences RIBS Okayama Japan Keywords phospholipase D phospholipid substrate recognition SPR Streptomyces Correspondence T. Hatanaka Research Institute for BiologicalSciences RIBS Okayama 7549-1 Kibichuo-cho Kaga-gun Okayama 716-1241 Japan Fax 81 866 56 9454 Tel 81 866 56 9452 E-mail hatanaka@ Received 12 January 2007 revised 14 March 2007 accepted 22 March 2007 Recently we identified Ala426 and Lys438 of phospholipase D from Strep-tomyces septatus TH-2 TH-2PLD as important residues for activity stability and selectivity in transphosphatidylation. These residues are located in a C-terminal flexible loop separate from two catalytic HxKxxxxD motifs. To study the role of these residues in substrate recognition we evaluated the affinities of inactive mutants in which these residues were substituted with Phe and His toward several phospholipids by SPR analysis. By substituting Ala426 and Lys438 with Phe and His respectively the inactive mutant showed a much stronger interaction with phosphatidylcholine and a weaker interaction with phosphatidylglycerol than the inactive TH-2PLD mutant. We demonstrated that Ala426 and Lys438 of TH-2PLD play a role in sensing the head group of phospholipids. doi Phospholipase D PLD EC catalyzes phospholipid hydrolysis and phosphatidyl transfer Fig. 1A . This is a ubiquitous and important enzyme involved in signal transduction in mammals 1 2 . Streptomyces PLDs can be categorized into two types one is an iron-containing enzyme such as that from Streptomyces chromofuscus chromofuscus PLD for which tightly bound iron is necessary for its catalytic activity 3 and the other is a member of the PLD superfamily whose hallmark is the possession of two catalytic HxKxxxxD HKD motifs 4-6 . Because enzymes of the latter type have a .

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