title: Sensor of phospholipids in Streptomyces phospholipase D.
abstract: 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.
authors: Research Institute for Biological Sciences (RIBS), Okayama, Japan.
authors: Yoshiko
authors: Y
authors: Uesugi
authors: Jiro
authors: J
authors: Arima
authors: Masaki
authors: M
authors: Iwabuchi
authors: Tadashi
authors: T
authors: Hatanaka
mesh_terms: D000595:Amino Acid Sequence; D008081:Liposomes / Q000378:metabolism; D010713:Phosphatidylcholines / Q000494:pharmacology; D010739:Phospholipase D / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D010743:Phospholipids / Q000378:metabolism* / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D013050:Spectrometry, Fluorescence; D013302:Streptomyces / Q000201:enzymology; D020349:Surface Plasmon Resonance
chemical_list: D008081:Liposomes; D010713:Phosphatidylcholines; D010743:Phospholipids; D010739:Phospholipase D; C028694:1-palmitoyl-2-oleoylphosphatidylcholine