Interaction of glucose transporter 1 with anion exchanger 1 in vitro.
The facilitative glucose transporter 1 (GLUT1) mediates the passive diffusion of d-glucose across the cell membrane, providing the energy resource in glycolysis in the erythrocytes. Anion exchanger 1 (band 3) is another important membrane protein that mediates rapid exchange of CO(2) through Cl(-)/HCO(3)(-) exchange across the erythrocyte membrane. For verifying the presumption over a decade that GLUT1 and band 3 in the erythrocyte would be interacting with each other, we cloned and expressed both the cytoplasmic domains of GLUT1 and band 3 in Escherichia coli, and tested their binding ability. By coimmunoprecipitation we found that among the tested N-terminal, C-terminal, and loop fraction of GLUT1, only the C-terminal of GLUT1 can interact with cytoplasmic domain of band 3. The interaction was further verified by coimmunoprecipitation and pull-down assay using both proteins as bait and target. These results showed that GLUT1 and band 3 form a protein complex that can regulate the activities of the proteins within it.
Jiang
Weihua
W
Department of Physiology and Biophysics, School of Life Sciences, Fudan University, Shanghai 200433, China.
Ding
Yu
Y
Su
Yang
Y
Jiang
Ming
M
Hu
Xiaojian
X
Zhang
Zhihong
Z
D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D051272:Glucose Transporter Type 1 / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011994:Recombinant Proteins / Q000378:metabolism
D001457:Anion Exchange Protein 1, Erythrocyte; D051272:Glucose Transporter Type 1; D011994:Recombinant Proteins