Phosphatidylinositol 3-phosphate induces the membrane penetration of the FYVE domains of Vps27p and Hrs.
The FYVE domain mediates the recruitment of proteins involved in membrane trafficking and cell signaling to phosphatidylinositol 3-phosphate (PtdIns(3)P)-containing membranes. To elucidate the mechanism by which the FYVE domain interacts with PtdIns(3)P-containing membranes, we measured the membrane binding of the FYVE domains of yeast Vps27p and Drosophila hepatocyte growth factor-regulated tyrosine kinase substrate and their mutants by surface plasmon resonance and monolayer penetration analyses. These measurements as well as electrostatic potential calculation show that PtdIns(3)P specifically induces the membrane penetration of the FYVE domains and increases their membrane residence time by decreasing the positive charge surrounding the hydrophobic tip of the domain and causing local conformational changes. Mutations of hydrophobic residues located close to the PtdIns(3)P-binding pocket or an Arg residue directly involved in PtdIns(3)P binding abrogated the penetration of the FYVE domains into the monolayer, the packing density of which is comparable with that of biological membranes and large unilamellar vesicles. Based on these results, we propose a mechanism of the membrane binding of the FYVE domain in which the domain first binds to the PtdIns(3)P-containing membrane by specific PtdIns(3)P binding and nonspecific electrostatic interactions, which is then followed by the PtdIns(3)P-induced partial membrane penetration of the domain.
Stahelin
Robert V
RV
Department of Chemistry, University of Illinois, Chicago, Illinois 60607, USA.
Long
Fei
F
Diraviyam
Karthikeyan
K
Bruzik
Karol S
KS
Murray
Diana
D
Cho
Wonhwa
W
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004330:Drosophila; D056827:Endosomal Sorting Complexes Required for Transport; D007700:Kinetics; D008869:Microtubule-Associated Proteins; D008958:Models, Molecular; D009154:Mutation; D018129:Phosphatidylinositol Phosphates / Q000235:genetics / Q000494:pharmacology*; D010750:Phosphoproteins / Q000378:metabolism*; D011487:Protein Conformation; D029701:Saccharomyces cerevisiae Proteins; D055672:Static Electricity; D014157:Transcription Factors / Q000378:metabolism*; D033921:Vesicular Transport Proteins; D016335:Zinc Fingers
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; C460141:FYCO1 protein, human; D008869:Microtubule-Associated Proteins; D018129:Phosphatidylinositol Phosphates; D010750:Phosphoproteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C119627:VPS27 protein, S cerevisiae; D033921:Vesicular Transport Proteins; C096164:hepatocyte growth factor-regulated tyrosine kinase substrate; C055525:phosphatidylinositol 3-phosphate