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Sensor of phospholipids in Streptomyces phospholipase D
Author(s) -
Uesugi Yoshiko,
Arima Jiro,
Iwabuchi Masaki,
Hatanaka Tadashi
Publication year - 2007
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2007.05802.x
Subject(s) - phosphatidylglycerol , phosphatidylcholine , mutant , chemistry , phospholipase , phospholipase d , stereochemistry , biochemistry , residue (chemistry) , affinities , streptomyces , substrate (aquarium) , phospholipid , enzyme , biology , gene , bacteria , genetics , ecology , membrane
Recently, we identified Ala426 and Lys438 of phospholipase D from Streptomyces 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.