Edelfosine is incorporated into rafts and alters their organization.
The effect of edelfosine (1- O-octadecyl-2- O-methyl-rac-glycero-3-phosphocholine or ET-18-OCH3) on model membranes containing 1-palmitoyl-2-oleoyl- sn-glycero-3-phosphocholine/sphingomyelin/cholesterol (POPC/SM/cholesterol) was studied by several physical techniques. The sample POPC/SM (1:1 molar ratio) showed a broad phase transition as seen by DSC, X-ray diffraction, and 2H NMR. The addition of edelfosine to this sample produced isotropic structures at temperatures above the phase transition, as seen by 2H NMR and by 31P NMR. When cholesterol was added to give a POPC/SM/cholesterol (at a molar ratio 1:1:1), no transition was observed by DSC nor X-ray diffraction, and 2H NMR indicated the presence of a liquid ordered phase. The addition of 10 mol % edelfosine increased the thickness of the membrane as seen by X-ray diffraction and led to bigger differences in the values of the molecular order of the membrane detected at high and low temperatures, as detected through the M 1 first spectral moment from 2H NMR. These differences were even greater when 20 mol % edelfosine was added, and a transition was now clearly visible by DSC. In addition, a gel phase was clearly indicated by X-ray diffraction at low temperatures. The same technique pointed to greater membrane thickness in this mixture and to the appearance of a second membrane structure, indicating the formation of two separated phases in the presence of edelfosine. All of these data strongly suggest that edelfosine associating with cholesterol alter the phase status present in a POPC/SM/cholesterol (1:1:1 molar ratio) mixture, which is reputed to be a model of a raft structure. However, cell experiments showed that edelfosine colocalizes in vivo with rafts and that it may reach concentrations higher than 20 mol % of total lipid, indicating that the concentrations used in the biophysical experiments were within what can be expected in a cell membrane. The conclusion is that molecular ways of action of edelfosine in cells may involve the modification of the structure of rafts.
Ausili
Alessio
A
Departamento de Bioquimica y Biologia Molecular A, Facultad de Veterinaria, Universidad de Murcia, Apartado de Correos 4021, Murcia, Spain.
Torrecillas
Alejandro
A
Aranda
Francisco J
FJ
Mollinedo
Faustino
F
Gajate
Consuelo
C
Corbalán-García
Senena
S
de Godos
Ana
A
Gómez-Fernández
Juan C
JC
D000970:Antineoplastic Agents / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry; D006801:Humans; D019169:Jurkat Cells; D009682:Magnetic Resonance Spectroscopy; D044367:Phase Transition; D010713:Phosphatidylcholines / Q000737:chemistry; D010726:Phosphodiesterase Inhibitors / Q000737:chemistry*; D010742:Phospholipid Ethers / Q000737:chemistry*; D013109:Sphingomyelins / Q000737:chemistry; D013696:Temperature; D014961:X-Ray Diffraction
D000970:Antineoplastic Agents; D010713:Phosphatidylcholines; D010726:Phosphodiesterase Inhibitors; D010742:Phospholipid Ethers; D013109:Sphingomyelins; C026659:edelfosine; D002784:Cholesterol