Molecular mechanism of membrane targeting by the GRP1 PH domain

Ju He, Rachel M. Haney, Mohsin Vora, Vladislav Verkhusha, Robert V. Stahelin, Tatiana G. Kutateladze

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

The general receptor for phosphoinositides isoform 1(GRP1) is recruited to the plasma membrane in response to activation of phosphoinositide 3-kinases and accumulation of phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P3]. GRP1's pleckstrin homology (PH) domain recognizes PtdIns(3,4,5)P3 with high specificity and affinity, however, the precise mechanism of its association with membranes remains unclear. Here, we detail the molecular basis of membrane anchoring by the GRP1 PH domain. Our data reveal a multivalent membrane docking involving PtdIns(3,4,5)P3 binding, regulated by pH and facilitated by electrostatic interactions with other anionic lipids. The specific recognition of PtdIns(3,4,5)P3 triggers insertion of the GRP1 PH domain into membranes. An acidic environment enhances PtdIns(3,4,5)P3 binding and increases membrane penetration as demonstrated by NMR and monolayer surface tension and surface plasmon resonance experiments. The GRP1 PH domain displays a 28 nM affinity for POPC/1-palmitoyl-2-oleoyl- sn-glycero-3-phosphoethanolamine/PtdIns(3,4,5)P3 vesicles at pH 6.0, but binds 22-fold weaker at pH 8.0. The pH sensitivity is attributed in part to the His355 residue, protonation of which is required for the robust interaction with PtdIns (3,4,5)P3 and significant membrane penetration, as illustrated by mutagenesis data. The binding affinity of the GRP1 PH domain for PtdIns(3,4,5)P3-containing vesicles is further amplified (by ∼6-fold) by nonspecific electrostatic interactions with phosphatidylserine/ phosphatidylinositol. Together, our results provide new insight into the multivalent mechanism of the membrane targeting and regulation of the GRP1 PH domain.

Original languageEnglish (US)
Pages (from-to)1807-1815
Number of pages9
JournalJournal of Lipid Research
Volume49
Issue number8
DOIs
StatePublished - Aug 2008

Fingerprint

Protein Isoforms
Membranes
Phosphatidylinositols
Coulomb interactions
Static Electricity
phosphatidylinositol receptors
platelet protein P47
phosphatidylinositol 3,4,5-triphosphate
Pleckstrin Homology Domains
Mutagenesis
Surface Tension
1-Phosphatidylinositol 4-Kinase
Surface Plasmon Resonance
Protonation
Phosphatidylserines
Surface plasmon resonance
Cell membranes
Surface tension
Monolayers
Phosphotransferases

Keywords

  • General receptor for phosphoinositides isoform 1
  • Phosphatidylinositol 3,4,5-trisphosphate
  • Phosphoinositide
  • Pleckstrin homology domain

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Endocrinology

Cite this

He, J., Haney, R. M., Vora, M., Verkhusha, V., Stahelin, R. V., & Kutateladze, T. G. (2008). Molecular mechanism of membrane targeting by the GRP1 PH domain. Journal of Lipid Research, 49(8), 1807-1815. https://doi.org/10.1194/jlr.M800150-JLR200

Molecular mechanism of membrane targeting by the GRP1 PH domain. / He, Ju; Haney, Rachel M.; Vora, Mohsin; Verkhusha, Vladislav; Stahelin, Robert V.; Kutateladze, Tatiana G.

In: Journal of Lipid Research, Vol. 49, No. 8, 08.2008, p. 1807-1815.

Research output: Contribution to journalArticle

He, J, Haney, RM, Vora, M, Verkhusha, V, Stahelin, RV & Kutateladze, TG 2008, 'Molecular mechanism of membrane targeting by the GRP1 PH domain', Journal of Lipid Research, vol. 49, no. 8, pp. 1807-1815. https://doi.org/10.1194/jlr.M800150-JLR200
He, Ju ; Haney, Rachel M. ; Vora, Mohsin ; Verkhusha, Vladislav ; Stahelin, Robert V. ; Kutateladze, Tatiana G. / Molecular mechanism of membrane targeting by the GRP1 PH domain. In: Journal of Lipid Research. 2008 ; Vol. 49, No. 8. pp. 1807-1815.
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