Human erythrocyte glucose transporter: normal asymmetric orientation and function in liposomes
| العنوان: | Human erythrocyte glucose transporter: normal asymmetric orientation and function in liposomes |
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| المؤلفون: | Loyal G. Tillotson, Susan Kalled, Chia-Chen Chen, Kurt J. Isselbacher, Tomonori Kurokawa, Shyh Yu Shaw |
| المصدر: | Proceedings of the National Academy of Sciences. 83:2652-2656 |
| بيانات النشر: | Proceedings of the National Academy of Sciences, 1986. |
| سنة النشر: | 1986 |
| مصطلحات موضوعية: | Monosaccharide Transport Proteins, Nucleoside Transport Proteins, Nucleoside transporter, Membrane Lipids, chemistry.chemical_compound, Escherichia coli, medicine, Humans, Band 3, Cytochalasin B, Octyl glucoside, Binding Sites, Multidisciplinary, biology, Chemistry, Erythrocyte Membrane, Glucose transporter, Membrane Proteins, Transporter, Blood Proteins, Membrane transport, Trypsin, Cytochalasins, Biochemistry, Liposomes, biology.protein, Research Article, medicine.drug |
| الوصف: | The transport function and orientation of the reconstituted human erythrocyte glucose transporter was studied with liposomes made with bovine brain lipid or Escherichia coli lipid. Reconstitution was achieved by a simple octyl glucoside dilution method. The reconstituted transporters with either lipid showed identical counterflow transport activity, the same response to various inhibitors, and characteristic cytochalasin B (CB) labeling. Functional location and purification of the glucose transporter was performed by using gel-permeation high-performance liquid chromatography with octyl glucoside-containing buffer. The reconstituted transport activity was associated only with band 4.5 protein (preactin) and not with band 3 protein. Both CB binding and transport function of the reconstituted transporters were resistant to trypsin but susceptible to chymotrypsin digestion. However, both trypsin and chymotrypsin treatment of unsealed ghosts completely eliminated the CB labeling and transport function of the glucose transporter. In our reconstitution system the glucose transporters maintained a normal asymmetrical (right-side-out) orientation and good transport function. A specific monoclonal antibody against the glucose transporter inhibited CB labeling of the transporters on unsealed ghosts. This was not found with the reconstituted system; however, after freeze-thawing there was a significant inhibition of CB binding by the antibody. These findings suggest that the CB-binding site of the reconstituted transporter is on the inner side of the proteoliposomes. |
| تدمد: | 1091-6490 0027-8424 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::dbd31597f257d651d472b9f3fac12145 https://doi.org/10.1073/pnas.83.8.2652 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....dbd31597f257d651d472b9f3fac12145 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 00278424 |
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