Behavior of a GPI-anchored protein in phospholipid monolayers at the air-water interface.
The interaction between alkaline phosphatase (AP), a glycosylphosphatidylinositol (GPI)-anchored protein (AP-GPI), and phospholipids was monitored using Langmuir isotherms and PM-IRRAS spectroscopy. AP-GPI was injected under C16 phospholipid monolayers with either a neutral polar head (1,2-dipalmitoyl-sn-glycero-3-phosphocholine monohydrate (DPPC)) or an anionic polar head (1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS)). The increase in molecular area due to the injection of protein depended on the surface pressure and the type of phospholipid. At all surface pressures, it was highest in the case of DPPS monolayers. The surface elasticity coefficient E, determined from the pi-A diagrams, allowed to deduct that the AP-GPI-phospholipid mixtures presented a molecular arrangement less condensed than the corresponding pure phospholipid films. PM-IRRAS spectra suggested different protein-lipid interactions as a function of the nature of the lipids. AP-GPI modified the organization of the DPPS deuterated chains whereas AP-GPI affected only the polar group of DPPC at low surface pressure (8 mN/m). Different protein hydration layers between the DPPC and DPPS monolayers were suggested to explain these results. PM-IRRAS spectra of AP-GPI in the presence of lipids showed a shape similar to those collected for pure AP-GPI, indicating a similar orientation of AP-GPI in the presence or absence of phospholipids, where the active sites of the enzyme are turned outside of the membrane.
Ronzon
Frédéric
F
Laboratoire de Physico-Chimie Biologique, UMR 5013, Université Claude Bernard Lyon I, Villeurbanne, France. ronzon@univ-lyon1.fr
Desbat
Bernard
B
Chauvet
Jean Paul
JP
Roux
Bernard
B
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry; D000469:Alkaline Phosphatase / Q000737:chemistry*; D001665:Binding Sites; D004548:Elasticity; D017261:Glycosylphosphatidylinositols / Q000737:chemistry*; D008567:Membranes, Artificial; D008968:Molecular Conformation; D010718:Phosphatidylserines / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry*; D011312:Pressure; D013055:Spectrophotometry, Infrared / Q000379:methods; D013499:Surface Properties; D013816:Thermodynamics
D017261:Glycosylphosphatidylinositols; D008567:Membranes, Artificial; D010718:Phosphatidylserines; D010743:Phospholipids; D015060:1,2-Dipalmitoylphosphatidylcholine; C038694:dipalmitoylphosphatidylserine; D000469:Alkaline Phosphatase