The Pleckstrin Homology Domain of Phospholipase C-δ1 Binds with High Affinity to Phosphatidylinositol 4,5-Bisphosphate in Bilayer Membranes

Pilar Garcia, Rakesh Gupta, Shefali Shah, Andrew J. Morris, Simon A. Rudge, Suzanne Scarlata, Victoria Petrova, Stuart McLaughlin, Mario J. Rebecchi

Research output: Contribution to journalArticlepeer-review

249 Scopus citations

Abstract

The pleckstrin homology (PH) domain of phospholipase C-δ1 (PLC-δ1) binds to phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) in phospholipid membranes with an affinity (Ka ~ 106 M-1) and specificity comparable to those of the native enzyme. PLC-δ1 and its PH domain also bind inositol 1,4,5-trisphosphate, the polar head group of PI(4,5)P2, with comparable affinity and approximately 1:1 stoichiometry. A peptide corresponding to amino acids 30-43 of the PLC-δ1 PH domain contains several basic residues predicted to bind PI(4,5)P2, but binds weakly and with little specificity for PI(4,5)P2; hence the tertiary structure of the isolated PH domain is required for high affinity PI(4,5)P2 binding. Our PI-(4,5)P2 binding results support the hypothesis that the intact PH domain, serving as a specific tether, directs PLC-δ1 to membranes enriched in PI(4,5)P2 and permits the active site, located elsewhere in the protein, to hydrolyze multiple substrate molecules before this enzyme dissociates from the membrane surface.

Original languageEnglish
Pages (from-to)16228-16234
Number of pages7
JournalBiochemistry
Volume34
Issue number49
DOIs
StatePublished - Dec 1995

ASJC Scopus subject areas

  • Biochemistry

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