pH-dependent domain formation in phosphatidylinositol polyphosphate/ phosphatidylcholine mixed vesicles

Abstract

Phosphatidylinositol polyphosphates (PI-PPs) have been shown to mediate a large variety of physiological processes by attracting proteins to specific cellular sites. Such site-specific signaling requires local accumulation of PI-PPs, and in light of the rich headgroup functionality, it is conceivable that hydrogen bond formation between adjacent headgroups is a contributing factor to the formation of PI-PP-enriched domains. To explore the significance of hydrogen bond formation for the mutual interaction of PI-PPs, this study aims to characterize the pH-dependent phase behavior of phosphatidylcholine/ phosphatidylinositol bisphosphate and trisphosphate mixed vesicles by differential scanning calorimetry, infrared transmission spectroscopy, and fluorescence resonance energy transfer measurements. For pH values >7-7.5, the experiments yielded results consistent with dipalmitoylphosphatidylcholine/ dipalmitoylphosphatidylinositol polyphosphate gel phase demixing, whereas for moderately acidic conditions, an enhanced mixing was observed. Similarly, this pH-dependent formation of PI-PP-enriched domains was also found for the physiologically important fluid phase. The stability of PI-PP-enriched domains and to some extent the pH dependence of the domain formation was governed by the number as well as the position of the phosphomonoester groups at the inositol ring. Copyright © 2005 by the American Society for Biochemistry and Molecular Biology, Inc.