A METHOD FOR UNIDIRECTIONAL RECONSTITUTION OF HUMAN ERYTHROCYTE GLUCOSE-TRANSPORTER

The glucose transporter of human erythrocytes (Glut1) was reconstitute d into soybean phospholipid liposomes by a method of direct incorporat ion using the nonionic detergent n-octyl beta-D-glucopyranoside. The r econstituted proteoliposomes were proved to be intact and low ionic pe rmeability. Freeze-fracture electron microscopy study showed that the diameter of the proteoliposomes was about 150 +/- 50 nm and the protei n was randomly distributed. The kinetic parameters of the reconstitute d transporter were: K-m = 16.23 mmol/L, V-max = 34.48 nmol/sec.mg prot ein. Furthermore, about 90% of the glucose transporter in the reconsti tuted proteoliposomes were orientated inside-out. Until now it is a mo re efficient method for uni-directional reconstitution of Glut1 with g ood reproducibility and higher transport activity.