Use of spin-labeled and fluorescent lipids to study the activity of the phospholipid transfer protein from maize seedlings.
The transfer of spin-labeled and fluorescent lipids between sonicated vesicles and different host membranes has been measured in the presence or absence of a phospholipid transfer protein purified from maize seedlings. It was found that the protein has little specificity towards the phospholipid head group and allows the transfer of hydrophobic long chain phospholipids. By contrast, no transfer of a cholesterol analogue could be detected. By EPR spectroscopy, evidence is presented that shows that the protein catalyzes the incorporation of labeled phospholipids in the outer monolayer of the acceptor membranes. The efficiency of the transfer depends largely on the nature of the acceptor: erythrocytes are more difficult to label than chromaffin granules or liposomes made with unsaturated lipids. Thus, consistent with the high activation energy observed, the transfer is facilitated when it involves fluid membranes. These results are in favor of a process involving the exchange of phospholipids, facilitated by a shuttle protein rather than a fusion mechanism.
Geldwerth
D
D
Institut de Biologie Physico-Chimique, Paris, France.
de Kermel
A
A
Zachowski
A
A
Guerbette
F
F
Kader
J C
JC
Henry
J P
JP
Devaux
P F
PF
D001205:Ascorbic Acid / Q000737:chemistry; D002352:Carrier Proteins / Q000378:metabolism*; D002837:Chromaffin Granules / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D004912:Erythrocytes / Q000737:chemistry; D005456:Fluorescent Dyes; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry; D008565:Membrane Proteins / Q000378:metabolism*; D049294:Phospholipid Transfer Proteins; D013113:Spin Labels; D013696:Temperature; D003313:Zea mays
D002352:Carrier Proteins; D005456:Fluorescent Dyes; D008055:Lipids; D008081:Liposomes; D008565:Membrane Proteins; D049294:Phospholipid Transfer Proteins; D013113:Spin Labels; D001205:Ascorbic Acid