| 摘要(英) |
Pmp1p is a small single-membrane span proteolipid that functions as a regulatory subunit of the plasma membrane H+-ATPase Pma1p. To explore the interaction of Pmp1p and lipids, we study the properties of GUV (giant unilamellar vesicle) model membranes composed of DPPC, POPC, ergosterol and protein Pmp1p by fluorescence and confocal microscopy. The fluorescence images of GUV membranes were taken as a function of temperature and Pmp1p concentration. The morphology of the GUV membranes is observed.
Addition of Pmp1p to 1:1:1 DPPC/POPC/ergosterol influences the morphology of GUV membranes. The bright phase (presumably in the ld phase) increases, and the dark phase (presumably in the lo phase) decreases with increasing Pmp1p concentration up to 0.2 mol% Pmp1p. This suggests that Pmp1p decreases the fraction of the lo-phase regions, ie. lipid rafts. In addition, the decrease of Tmix also indicates that Pmp1p decreases the order of GUV membranes. It is found that Pmp1p locates in the bright-phase regions, implying that Pmp1p is preferentially associated with the non-raft region rather than the raft region. This is in disagreement with current views. It is believed that Pmp1p regulates the function of another raft protein H+-ATPase (Pma1p). Our results suggest that proteins Pmp1p and Pma1p exist in different parts of the membrane, and Pmp1p interacts with Pma1p indirectly via an unknown mechanism.
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