Mechanism of vitamin A movement between rod outer segments, interphotoreceptor retinoid-binding protein, and liposomes.
Vitamin A movement between rod outer segment (ROS) membranes, interphotoreceptor retinoid-binding protein (IRBP), and liposomes was examined by two different methods. Equilibrium exchange of all-trans-retinol was followed by assessing the transfer of [3H]retinol from liposomes to ROS membranes as compared to a nontransferable marker, [14C]triolein. In the absence of IRBP, a rapid, spontaneous transfer of [3H] retinol to the ROS membranes occurred. In the presence of 2 microM IRBP, retinol transfer decreased by approximately one-half, whereas a similar concentration of bovine serum albumin had no effect on this spontaneous transfer. Kinetics of retinol transfer between single unilamellar vesicles were determined by the method of fluorescence energy transfer. The first order rate constant for this transfer was 0.85 s-1 at 22 degrees C at either pH 7.4 or pH 2.8. This rate was not affected by varying the concentration of acceptor vesicles 50-fold or by varying their concentration 10-fold at a constant ratio of donor-to-acceptor vesicles. The presence of IRBP as an additional acceptor did not change the rate. The transfer was temperature-dependent with an activation energy of 7.8 kcal/mol. The transfer rate appeared to be an increasing exponential function of ionic strength since high concentrations of NaCl decreased the transfer rate significantly. The transfer rate of retinol from IRBP to single unilamellar vesicles also followed first order kinetics with a rate constant of 0.11 s-1 at 22 degrees C, which was approximately 8 times slower than that of transfer between vesicles. We conclude that the transfer of all-trans-retinol between liposomes and membranes can be accomplished rapidly via the aqueous phase, and that IRBP retards rather than facilitates this transfer process.
Ho
M T
MT
Program in Neuroscience, Baylor College of Medicine, Houston, Texas 77030.
Massey
J B
JB
Pownall
H J
HJ
Anderson
R E
RE
Hollyfield
J G
JG
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism*; D010714:Phosphatidylethanolamines / Q000378:metabolism*; D010786:Photoreceptor Cells / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism*; D014304:Triolein / Q000378:metabolism; D014801:Vitamin A / Q000378:metabolism*
D008081:Liposomes; D010169:Palmitic Acids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D012177:Retinol-Binding Proteins; D014801:Vitamin A; D014304:Triolein; D019308:Palmitic Acid