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Phosphatidylserine induces functional and structural alterations of the membrane‐associated pleckstrin homology domain of phospholipase C‐δ1
Author(s) -
Uekama Naoko,
Sugita Takio,
Okada Masashi,
Yagisawa Hitoshi,
Tuzi Satoru
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.1742-4658.2006.05574.x
Subject(s) - pleckstrin homology domain , chemistry , c2 domain , vesicle , phosphatidylserine , membrane , phosphatidylinositol , crystallography , biophysics , biochemistry , phospholipid , biology , signal transduction
The membrane binding affinity of the pleckstrin homology (PH) domain of phospholipase C (PLC)‐δ1 was investigated using a vesicle coprecipitation assay and the structure of the membrane‐associated PH domain was probed using solid‐state 13 C NMR spectroscopy. Twenty per cent phosphatidylserine (PS) in the membrane caused a moderate but significant reduction of the membrane binding affinity of the PH domain despite the predicted electrostatic attraction between the PH domain and the head groups of PS. Solid‐state NMR spectra of the PH domain bound to the phosphatidylcholine (PC)/PS/phosphatidylinositol 4,5‐bisphosphate (PIP 2 ) (75 : 20 : 5) vesicle indicated loss of the interaction between the amphipathic α2‐helix of the PH domain and the interface region of the membrane which was previously reported for the PH domain bound to PC/PIP 2 (95 : 5) vesicles. Characteristic local conformations in the vicinity of Ala88 and Ala112 induced by the hydrophobic interaction between the α2‐helix and the membrane interface were lost in the structure of the PH domain at the surface of the PC/PS/PIP 2 vesicle, and consequently the structure becomes identical to the solution structure of the PH domain bound to d ‐ myo ‐inositol 1,4,5‐trisphosphate. These local structural changes reduce the membrane binding affinity of the PH domain. The effects of PS on the PH domain were reversed by NaCl and MgCl 2 , suggesting that the effects are caused by electrostatic interaction between the protein and PS. These results generally suggest that the structure and function relationships among PLCs and other peripheral membrane proteins that have similar PH domains would be affected by the local lipid composition of membranes.