Association of sterol- and glycosylphosphatidylinositol-linked proteins with Drosophila raft lipid microdomains.
In vertebrates, the formation of raft lipid microdomains plays an important part in both polarized protein sorting and signal transduction. To establish a system in which raft-dependent processes could be studied genetically, we have analyzed the protein and lipid composition of these microdomains in Drosophila melanogaster. Using mass spectrometry, we identified the phospholipids, sphingolipids, and sterols present in Drosophila membranes. Despite chemical differences between Drosophila and mammalian lipids, their structure suggests that the biophysical properties that allow raft formation have been preserved. Consistent with this, we have identified a detergent-insoluble fraction of Drosophila membranes that, like mammalian rafts, is rich in sterol, sphingolipids, and glycosylphosphatidylinositol-linked proteins. We show that the sterol-linked Hedgehog N-terminal fragment associates specifically with this detergent-insoluble membrane fraction. Our findings demonstrate that raft formation is preserved across widely separated phyla in organisms with different lipid structures. They further suggest sterol modification as a novel mechanism for targeting proteins to raft membranes and raise the possibility that signaling and polarized intracellular transport of Hedgehog are based on raft association.
Rietveld
A
A
Cell Biology Programme, European Molecular Biology Laboratory, Meyerhofstrasse-1, 69117 Heidelberg, Germany.
Neutz
S
S
Simons
K
K
Eaton
S
S
D000818:Animals; D003902:Detergents; D004330:Drosophila; D029721:Drosophila Proteins; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D053823:Hedgehog Proteins; D019476:Insect Proteins / Q000378:metabolism; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D015394:Molecular Structure; D021382:Protein Sorting Signals / Q000378:metabolism; D013261:Sterols / Q000378:metabolism*
D003902:Detergents; D029721:Drosophila Proteins; D017261:Glycosylphosphatidylinositols; D053823:Hedgehog Proteins; D019476:Insect Proteins; D008563:Membrane Lipids; D008565:Membrane Proteins; D021382:Protein Sorting Signals; D013261:Sterols; C077245:hh protein, Drosophila