Asymmetric transport of a fluorescent glucose analogue by human erythrocytes

Speizer, L.A.; Haugland, Richard P.; Kutchai, Howard

doi:10.1016/0005-2736(85)90476-6

Cited by 94 publications

(92 citation statements)

References 39 publications

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“…The total amount of radioactivity taken up by cells or ghosts was measured by means of a scintillation counter (Packard Tri-Carb 2000 CA anatyser, Packard Instrument Co, Downers Grove, IL, USA), using 0.5ml of 10% (w/v) trichloroacetic acid extracts obtained by centrifugation. Uptake of [3H]T was measured in pmol//d of intracellular water, which was determined according to Speizer et aL (1985).…”

Section: Methodsmentioning

confidence: 99%

Thiamine transport by erythrocytes and ghosts in thiamine‐responsive megaloblastic anaemia

Rindi

Casirola

Poggi

et al. 1991

J of Inher Metab Disea

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A 9-year study of thiamine metabolism and cellular transport was performed in two patients with thiamine-responsive megaloblastic anaemia associated with diabetes mellitus and sensorineural deafness, in their relatives, and in age-matched controls from the same area. The ratios between the content of thiamine and that of its phosphoesters in erythrocytes were within the normal range, whereas the absolute values of thiamine and thiamine compounds were reduced by about 40% as compared to controls. Thiamine pyrophosphokinase activity was about 30% lower than in controls. Thiamine treatment restored the levels of thiamine and thiamine compounds to normal values, whereas kinase was unaffected. Both the saturable (specific, predominant at low, less than 2 mumol/L, physiological concentrations of thiamine) and the non-saturable component of thiamine transport were investigated. Erythrocytes and ghosts from patients exhibited no saturable component, this abnormality being specific for the patients and not shared by their parents. It is concluded that the cells from thiamine-responsive megaloblastic anaemia patients contain low levels of thiamine compounds, probably due to their inability to take up and retain physiological concentrations of thiamine, as a result of the lack of the saturable, specific component of transport and reduced thiamine pyrophosphokinase.

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Section: Methodsmentioning

confidence: 99%

Thiamine transport by erythrocytes and ghosts in thiamine‐responsive megaloblastic anaemia

Rindi

Casirola

Poggi

et al. 1991

J of Inher Metab Disea

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“…It has already been shown that it is transported through the cell membrane using the same mechanism as glucose, and that it is a substrate for hexokinase (19)(20)(21). Although transported by the same mechanism, 6-NBDG has a fluorophore at the phosphorylation site and is, therefore, not a substrate for hexokinase (22). Cy5.5 dye has better spectral characteristics than NBD, emitting light in the NIR region where there is minimal spectral interference from biological tissues (23).…”

Section: Introductionmentioning

confidence: 99%

Fluorescent Fructose Derivatives for Imaging Breast Cancer Cells

et al. 2007

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Breast cancer cells are known to overexpress Glut5, a sugar transporter responsible for the transfer of fructose across the cell membrane. Since Glut5 transporter is not significantly expressed in normal breast cells, fructose uptake can potentially be used to differentiate between normal and cancerous cells. Fructose was labeled with two fluorophores at the C-1 position: 7-nitro-1,2,3-benzadiazole (NBD) and Cy5.5. The labeling site was chosen on the basis of the presence and substrate specificity of the key proteins involved in the first steps of fructose metabolism. Using fluorescence microscopy, the uptake of the probes was studied in three breast cancer cell lines: MCF 7, MDA-MB-435, and MDA-MB-231. Both fluorescent fructose derivatives showed a very good uptake in all tested cell lines. The level of uptake was comparable to that of the corresponding glucose analogs, 2-NBDG and Cy5.5-DG. Significant uptake of 1-NBDF derivative was not observed in cells lacking Glut5 transporter, while the uptake of the 1-Cy5.5-DF derivative was independent of the presence of a fructose-specific transporter. While 1-NBDF showed Glut5-specific accumulation, the coupling of a large fluorophore such as Cy5.5 likely introduces big structural and electronic changes, leading to a fructose derivative that does not accurately describe the uptake of fructose in cells.

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“…reported a fluorescent derivative of glucose, 6-NBDG. 7 ) They found that 6-NBDG was incorporated in human erythrocytes and that its incorporation was competitively inhibited by D-glucose. That suggested the involvement of a glucose transporter in the erythrocyte membrane.…”

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confidence: 99%

“…That contrasted with the case of 6-NBDG, which was not modified in the cells. 7) To use 2-NBDG as a viability indicator, it is important to elucidate the intracellular fate of 2-NBDG. As the first step, are tried to clarify what is this fluorescent derivative of 2-NBDG found in E. coli cells immediately after incorporation.…”

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confidence: 99%