Cholate-solubilized erythrocyte glucose transporters exist as a mixture of homodimers and homotetramers
dc.contributor.author | Hebert, Daniel N | |
dc.contributor.author | Carruthers, Anthony | |
dc.date | 2022-08-11T08:08:58.000 | |
dc.date.accessioned | 2022-08-23T16:14:21Z | |
dc.date.available | 2022-08-23T16:14:21Z | |
dc.date.issued | 1991-05-14 | |
dc.date.submitted | 2008-03-21 | |
dc.identifier.citation | <p>Biochemistry. 1991 May 14;30(19):4654-8.</p> | |
dc.identifier.issn | 0006-2960 (Print) | |
dc.identifier.doi | 10.1021/bi00233a003 | |
dc.identifier.pmid | 2029513 | |
dc.identifier.uri | http://hdl.handle.net/20.500.14038/33841 | |
dc.description.abstract | The molecular size of purified, human erythrocyte glucose transport protein (GLUT1) solubilized in cholic acid was determined by size-exclusion chromatography (SEC) and sucrose gradient ultracentrifugation. GLUT1 purified in the presence of dithiothreitol (GLUT1 + DTT) is resolved as a complex of average Stokes' radius 5.74 nm by SEC. This complex displays D-glucose-inhibitable cytochalasin B binding and, upon reconstitution into proteoliposomes, catalyzes cytochalasin B inhibitable D-glucose transport. GLUT1 purified in the absence of dithiothreitol (GLUT1-DTT) is resolved by SEC as at least two particles of average Stokes' radii 5.74 (minor component) and 7.48 nm (major component). Solubilization of GLUT1-DTT in the presence of dithiothreitol reduces the amount of 7.48-nm complex and increases the amount of 5.74-nm complex resolved by SEC. GLUT1-DTT displays D-glucose-inhibitable cytochalasin B binding and, upon reconstitution into proteoliposomes, catalyzes cytochalasin B inhibitable D-glucose transport. Sucrose gradient ultracentrifugation of GLUT1 + DTT in cholate resolves GLUT1 into two components of 4.8 and 7.6 S. The 4.8S complex is the major component of GLUT1 + DTT. The reverse profile is observed upon sucrose gradient ultracentrifugation of GLUT1-DTT. SEC of human erythrocyte membrane proteins resolves GLUT1 as a major broad peak of average Stokes' radius 7.48 nm and a minor component of 5.74 nm. Both components are characterized by D-glucose-inhibitable cytochalasin B binding. Purified GLUT1 is associated with approximately 26 tightly bound lipid molecules per monomer of transport protein. These data suggest that purified GLUT1 exists as a mixture of homodimers and homotetramers in cholate-lipid micelles and that the presence of reductant during solubilization favors dimer formation. | |
dc.language.iso | en_US | |
dc.relation | <p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=2029513&dopt=Abstract ">Link to article in PubMed</a></p> | |
dc.relation.url | https://doi.org/10.1021/bi00233a003 | |
dc.subject | Catalysis; Cholic Acids; Chromatography, Gel; Cytochalasin B; Dithiothreitol; Enzyme-Linked Immunosorbent Assay; Erythrocyte Membrane; Erythrocytes; Humans; Membrane Proteins; Monosaccharide Transport Proteins; Ultracentrifugation | |
dc.subject | Life Sciences | |
dc.subject | Medicine and Health Sciences | |
dc.title | Cholate-solubilized erythrocyte glucose transporters exist as a mixture of homodimers and homotetramers | |
dc.type | Journal Article | |
dc.source.journaltitle | Biochemistry | |
dc.source.volume | 30 | |
dc.source.issue | 19 | |
dc.identifier.legacycoverpage | https://escholarship.umassmed.edu/gsbs_sp/5 | |
dc.identifier.contextkey | 467855 | |
html.description.abstract | <p>The molecular size of purified, human erythrocyte glucose transport protein (GLUT1) solubilized in cholic acid was determined by size-exclusion chromatography (SEC) and sucrose gradient ultracentrifugation. GLUT1 purified in the presence of dithiothreitol (GLUT1 + DTT) is resolved as a complex of average Stokes' radius 5.74 nm by SEC. This complex displays D-glucose-inhibitable cytochalasin B binding and, upon reconstitution into proteoliposomes, catalyzes cytochalasin B inhibitable D-glucose transport. GLUT1 purified in the absence of dithiothreitol (GLUT1-DTT) is resolved by SEC as at least two particles of average Stokes' radii 5.74 (minor component) and 7.48 nm (major component). Solubilization of GLUT1-DTT in the presence of dithiothreitol reduces the amount of 7.48-nm complex and increases the amount of 5.74-nm complex resolved by SEC. GLUT1-DTT displays D-glucose-inhibitable cytochalasin B binding and, upon reconstitution into proteoliposomes, catalyzes cytochalasin B inhibitable D-glucose transport. Sucrose gradient ultracentrifugation of GLUT1 + DTT in cholate resolves GLUT1 into two components of 4.8 and 7.6 S. The 4.8S complex is the major component of GLUT1 + DTT. The reverse profile is observed upon sucrose gradient ultracentrifugation of GLUT1-DTT. SEC of human erythrocyte membrane proteins resolves GLUT1 as a major broad peak of average Stokes' radius 7.48 nm and a minor component of 5.74 nm. Both components are characterized by D-glucose-inhibitable cytochalasin B binding. Purified GLUT1 is associated with approximately 26 tightly bound lipid molecules per monomer of transport protein. These data suggest that purified GLUT1 exists as a mixture of homodimers and homotetramers in cholate-lipid micelles and that the presence of reductant during solubilization favors dimer formation.</p> | |
dc.identifier.submissionpath | gsbs_sp/5 | |
dc.contributor.department | Department of Biochemistry and Molecular Pharmacology | |
dc.contributor.department | Graduate School of Biomedical Sciences | |
dc.source.pages | 4654-8 |