Biophysical and computational studies of membrane penetration by the GRP1 pleckstrin homology domain

Craig N. Lumb, Ju He, Yi Xue, Phillip J. Stansfeld, Robert V. Stahelin, Tatiana G. Kutateladze, Mark S.P. Sansom

Research output: Contribution to journalArticle

46 Scopus citations

Abstract

The pleckstrin homology (PH) domain of the general receptor for phosphoinositides 1 (GRP1) exhibits specific, high-affinity, reversible binding to phosphatidylinositol (3,4,5)-trisphosphate (PI(3,4,5)P 3) at the plasma membrane, but the nature and extent of the interaction between this bound complex and the surrounding membrane environment remains unclear. Combining equilibrium and nonequilibrium molecular dynamics (MD) simulations, NMR spectroscopy, and monolayer penetration experiments, we characterize the membrane-associated state of GRP1-PH. MD simulations show loops flanking the binding site supplement the interaction with PI(3,4,5)P 3 through multiple contacts with the lipid bilayer. NMR data show large perturbations in chemical shift for these loop regions on binding to PI(3,4,5)P 3-containing DPC micelles. Monolayer penetration experiments and further MD simulations demonstrate that mutating hydrophobic residues to polar residues in the flanking loops reduces membrane penetration. This supports a "dual-recognition" model of binding, with specific GRP1-PH-PI(3,4,5)P 3 interactions supplemented by interactions of loop regions with the lipid bilayer.

Original languageEnglish (US)
Pages (from-to)1338-1346
Number of pages9
JournalStructure
Volume19
Issue number9
DOIs
StatePublished - Sep 7 2011

    Fingerprint

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Lumb, C. N., He, J., Xue, Y., Stansfeld, P. J., Stahelin, R. V., Kutateladze, T. G., & Sansom, M. S. P. (2011). Biophysical and computational studies of membrane penetration by the GRP1 pleckstrin homology domain. Structure, 19(9), 1338-1346. https://doi.org/10.1016/j.str.2011.04.010