A method for uni-directional reconstitution of human erythrocyte glucose transporter.
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.
Xiao
L
L
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing.
Xie
W S
WS
Yang
F Y
FY
D003902:Detergents / Q000737:chemistry* / Q000494:pharmacology; D004912:Erythrocytes / Q000378:metabolism*; D005614:Freeze Fracturing; D005947:Glucose / Q000378:metabolism / Q000493:pharmacokinetics; D051272:Glucose Transporter Type 1; D005960:Glucosides / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008081:Liposomes; D008854:Microscopy, Electron; D009004:Monosaccharide Transport Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D010743:Phospholipids / Q000737:chemistry; D011510:Proteolipids / Q000737:chemistry
D003902:Detergents; D051272:Glucose Transporter Type 1; D005960:Glucosides; D008081:Liposomes; D009004:Monosaccharide Transport Proteins; D010743:Phospholipids; D011510:Proteolipids; C495677:SLC2A1 protein, human; C018619:octyl-beta-D-glucoside; D005947:Glucose