A method for uni-directional reconstitution of human erythrocyte glucose transporter.

Abstract

The glucose transporter of human erythrocytes (Glut1) was reconstituted into soybean phospholipid liposomes by a method of direct incorporation using the nonionic detergent n-octyl beta-D-glucopyranoside. The reconstituted proteoliposomes were proved to be intact and low ionic permeability. Freeze-fracture electron microscopy study showed that the diameter of the proteoliposomes was about 150 +/- 50 nm and the protein was randomly distributed. The kinetic parameters of the reconstituted transporter were: Km =16.23 mmol/L, Vmax = 34.48 nmol/sec x mg protein. Furthermore, about 90% of the glucose transporter in the reconstituted proteoliposomes were orientated inside-out. Until now it is a more efficient method for uni-directional reconstitution of Glut1 with good reproducibility and higher transport activity.