An Investigation of the Nano-Organization of Glucose Transporters, Glut1 and Glut3, in the Mammalian Plasma Membrane

• Main Author: Sireesha, Dommaraju
• Format: Others
• Language: English
• Published: Högskolan i Skövde, Institutionen för vård och natur 2008
• Subjects: Sireesha; Dommaraju; Gluts; Sweden; Denmark; skovde; Odense; skovde University; SDU university; Molecular biology; Molekylärbiologi
• Online Access: http://urn.kb.se/resolve?urn=urn:nbn:se:his:diva-2330

Glucose is a monosaccharide and fuel for body, it cannot pass through membrane by simple diffusion so, integral transmembrane proteins named glucose transporters (Gluts) are involved in the regulation of the movement of glucose between the extracellular and intracellular spaces within the body. GLUT1 and GLUT3 have previously been shown by cold detergent extraction methods to reside in distinct plasma membrane domains in non-polarized mammalian cells, with GLUT1, but not GLUT3, residing  in detergent-resistant membrane (DRM) domains. To confirm this observation under less invasive conditions, molecular fusion tags are inserted in the first external loop in Glut1 with biotin ligase acceptor peptide (BLAP) between Ser-55 and Ile-56 and in Glut3 with Acyl carrier peptide (ACP) in between Val-57 and Leu-58 respectively. These Glut fusion proteins will be used in order to confirm these observations by fluorescence recovery after photobleaching (FRAP) and single molecule fluorescence microscopy in live cells. hGLUT1-EGFP, hGLUT1 (AgeI)-EGFP recombinants were constructed and transfected human embryonic kidney cells (HEK-293) quantum dot images supports the fact that EGFP transfected cells uniformly and is distributed in the cell cytoplasm, hGLUT1-EGFP transfected cells and is localized to the cell membrane and hGLUT1 (AgeI)-EGFP transfected cells and located to the plasma membrane with high intensity.