Model system for heat-induced translocation of cytoplasmic beta-galactosidase across phospholipid bilayer membrane.
The possibility of the translocation of the enzyme across the phospholipid bilayer membrane was investigated by using the liposomes prepared by 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) in which beta-galactosidase (beta-gal) was entrapped. Exposing the POPC liposomes entrapping beta-gal inside to heat treatment (40-50 degrees C, 1-60 min) was found to induce its translocation across the liposome membrane. The translocated activity of beta-gal from inner to outer aqueous phase of liposomes indicated the maximal value when the liposomes entrapping beta-gal were heated at 45 degrees C for 30 min. The gel permeation profiles of the liposomes before and after heat treatment (45 degrees C, 30 min) also supported the translocation of beta-gal across the liposome membrane. The membrane fluidity of liposomes was found to be increased with increasing temperature, so that the hydrophobicity of liposome membrane was also increased. The local hydrophobicity of beta-gal was maximized at the temperature of 40-50 degrees C. The mechanisms of beta-gal translocation have been suggested to be triggered by the enhancement of hydrophobic interaction between the liposome surface and beta-gal molecules. Finally, a minimal scheme of possible mechanism on the heat-induced translocation of beta-gal has been presented on the basis of the hydrophobic interaction between the liposome and the proteins. The experimental data on the heat-induced translocation of beta-gal were well corresponding to those from model calculation.
Umakoshi
H
H
Department of Chemical Science and Engineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyamacho, Toyonaka, Osaka 560-0043, Japan.
Yoshimoto
M
M
Shimanouchi
T
T
Kuboi
R
R
Komasawa
I
I
D002850:Chromatography, Gel; D003198:Computer Simulation; D003593:Cytoplasm / Q000201:enzymology*; D006358:Hot Temperature; D008051:Lipid Bilayers; D008081:Liposomes; D008560:Membrane Fluidity; D008954:Models, Biological; D010743:Phospholipids; D012995:Solubility; D013499:Surface Properties; D014867:Water / Q000737:chemistry; D001616:beta-Galactosidase / Q000378:metabolism*
D008051:Lipid Bilayers; D008081:Liposomes; D010743:Phospholipids; D014867:Water; D001616:beta-Galactosidase