Glycosylation of the human erythrocyte glucose transporter is essential for glucose transport activity

Feugeas, Jean-Paul; Néel, Dominique; Pavia, A. A.; Laham, A.; Goussault, Yves; Derappe, Christian

doi:10.1016/0005-2736(90)90238-j

Cited by 27 publications

(10 citation statements)

References 23 publications

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“…Therefore, at least for this serum, we cannot exclude that part of the inhibition is due to the binding of the antibodies to glycans. Yet, in the case of the human Na'-glucose cotransporter, which is a glycoprotein, it has been clearly shown that the glycan part is absolutely necessary for the transport function (8). The possibility that some of the antibodies contained in the rabbit anti-42 kD serum may be directed towards glycans of the 42 kD region glycoproteins therefore does not contradict the conclusion that a 42 kD polypeptide (possibly a glycoprotein) is involved in the active transport of sucrose.…”

Section: Rabbit Polyclonal Serummentioning

confidence: 83%

Selective Inhibition of Active Uptake of Sucrose into Plasma Membrane Vesicles by Polyclonal Sera Directed against a 42 Kilodalton Plasma Membrane Polypeptide

Gallet

Lemoine²,

Gaillard³

et al. 1992

Plant Physiol.

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Several polyclonal sera were raised in rabbits and in mice against putative sucrose carrier proteins, I.e. a 42 kilodalton (O Gallet, R Lemoine, C Larsson, S Delrot [1989] Biochim Biophys Acta 978: 56-64) and a 62 kD (KG Ripp, PV Viitanen, WD Hitz, VR Fransceschi [1988] Plant Physiol 88: 1435-1445) polypeptide of the plasma membrane. The effects of these sera on the active uptake of sucrose and of valine into purified plasma membrane vesicles from sugar beet (Beta vulgaris L.) leaves and roots were studied. At a dilution of 1/50, the anti-42 kilodalton sera consistently inhibited sucrose uptake in plasma membranes from leaves or from roots. They had no effect on valine uptake. Under the same experimental conditions, the anti-62 kilodalton sera had no effect on active uptake of sucrose. The data further support the view that a 42 kilodalton polypeptide is a component of the transport system mediating sucrose uptake across the plasma membrane of plant cells.According to the mass-flow mechanism of translocation, long distance transport of sugars in the plant depends on the build-up and on the maintenance of a pressure gradient between the phloem ofthe source and the phloem of the sink. In this model, accepted by most authors, the path is thought to play a relatively passive role, whereas phloem loading and unloading govern the intensity and the direction of phloem transport by controlling sucrose fluxes. Although recent work indicates the importance of the symplastic pathway in some steps of assimilate transport (18,(25)(26)(27)(28), transport of sugars across the plasmalemma is involved in the control of major processes such as assimilate leakage from the mesophyll, loading (or retrieval) of sucrose in the phloem, and sugar unloading in mature sinks. The carrier protein(s) mediating sucrose transport through the plasma membrane therefore play(s) a major physiological role for the distribution of carbohydrates, and indirectly for all the phloem-mobile solutes moved by mass-flow. Unfortunately, these proteins have not yet been identified unequivocally. A sucrose photolyzable derivative, 6'-deoxy-6'-(4-azido-2-hydroxy) benzamidosucrose was used to label a sucrose binding protein in microsomal preparations from soybean cotyledons (24). This polypeptide appeared in the microsomal fraction of soybean cotyledon cells concomitantly with the onset of active sucrose influx (24). Polyclonal antibodies raised against the 62 kD polypeptide from soybean cross-reacted with a 62 kD polypeptide in purified plasma membrane vesicles from spinach leaves and immunolocalization studies in fully expanded leaves of this species showed that this polypeptide is specifically located in the plasma membrane ofthe sieve tube. The sieve tube plasma membranes were not immunolabeled in young leaves where the phloem contained no mature sieve tube member (29). The same localization was found in mature leaves of sugar beet with the same antibodies, but no Western blot was provided to show what proteins of sugar beet cross-reacted with the ...

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Section: Rabbit Polyclonal Serummentioning

confidence: 83%

Selective Inhibition of Active Uptake of Sucrose into Plasma Membrane Vesicles by Polyclonal Sera Directed against a 42 Kilodalton Plasma Membrane Polypeptide

Gallet

Lemoine²,

Gaillard³

et al. 1992

Plant Physiol.

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“…The oligosaccharide chain on the glucose transporter has previously been suggested to play a role in maintaining the optimal transport capacity of GLUT1 (Asano et al 1991;Feugeas et al 1990). Immunoblot detection of GLUT1 in HEMPAS erythrocytes (Fig.…”

Section: Anion and Glucose Transport Functions In Hempas Erythrocytesmentioning

confidence: 97%

Structural and functional consequences of an N-glycosylation mutation (HEMPAS) affecting human erythrocyte membrane glycoproteins

Kameh¹,

Landolt-Marticorena²,

Charuk³

et al. 1998

Biochem. Cell Biol.

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Band 3, the human erythrocyte anion exchanger (AE1), and the glucose transporter (GLUT1) proteins each contain a single site of N-glycosylation that is heterogeneously glycosylated. Lectin binding and enzymatic deglycosylation assays showed that the polylactosaminyl oligosaccharide structure of these glycoproteins was altered to a high mannose or hybrid glycan form in three patients with hereditary erythroblastic multinuclearity, with a positive acidified-serum lysis test (HEMPAS). Offspring from one of the HEMPAS patients had intermediate levels of polylactosaminyl oligosaccharide associated with AE1 and GLUT1, suggesting they may have been heterozygous for the genetic defect. The array of polylactosaminyl-containing glycoproteins present in EBV-transformed lymphoblasts derived from fresh blood of HEMPAS patients was similar to control lymphoblasts. HEMPAS lymphoblasts do not therefore express the defect in polylactosamine synthesis found in erythroid cells, indicating that lymphoid cells are not deficient in the processing enzymes or contain an alternative oligosaccharide processing pathway. Purified HEMPAS band 3 had an unaltered oligomeric structure but dimers aggregated more rapidly in detergent solution than normal band 3. The altered oligosaccharide structure did not affect the sensitivity of band 3 to proteolytic digestion in intact red cells but a greater amount of HEMPAS band 3 was associated with the cytoskeleton. The transport activities of AE1 and GLUT1 in HEMPAS erythrocytes were similar to those in normal controls. This shows that the HEMPAS glycosylation defect does not impair the functional accumulation of these two important erythrocyte membrane transporters even though it produces subtle structural changes in band 3 that result in its increased cytoskeletal interaction and self association in detergent solution.

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“…Modifications of the oligosaccharide structure of glycoproteins related to changes in the biological activity of transporter proteins have been described. The GLUT1 glucose transporter (21,22), the organic cation transporter of renal brush-border membrane (23), are examples of such a correlation. However, glycosylation is not required for the transport activity of the serotonin transporter (24).…”

Section: Discussionmentioning

confidence: 99%

Heterologous Expression and Functional Characterization of a Mouse Renal Organic Anion Transporter in Mammalian Cells

Kuze

Graves

Leahy

et al. 1999

Journal of Biological Chemistry

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Organic anion transporters play an essential role in eliminating a wide range of organic anions including endogenous compounds, xenobiotics, and their metabolites from kidney, thereby preventing their potentially toxic effects within the body. The goal of this study was to extend our previous study on the functional characterization and posttranslational modification of a mouse kidney organic anion transporter (mOAT), in a mammalian cell system, COS-7 cells. The transporter-mediated p-aminohippurate (PAH) uptake was saturable, probenecid-sensitive, and inhibited by a wide range of organic anions including vitamins, antihypertensive drugs, anti-tumor drugs, and anti-inflammatory drugs. Tunicamycin, an inhibitor of asparagine-linked glycosylation, significantly inhibited the transport activity. Immunofluorescence provided evidence that most of the protein remained in the intracellular compartment in tunicamycin-treated cells. Diethyl pyrocarbonate (DEPC), a histidine residue-specific reagent, completely blocked PAH transport. The inhibitory effect by DEPC was significantly protected (90%) by pretreating the cells with excess unlabeled PAH, suggesting that the histidine residues may be close to the PAH binding sites. Finally, in situ mRNA localization was studied in postnatal mouse kidney. The expression was observed in proximal tubules throughout development. We conclude that COS-7 cells may be useful in pharmacological and molecular biological studies of this carrier. The carbohydrate moieties are necessary for the proper trafficking of mOAT to the plasma membrane, and histidine residues appear to be important for the transport function.

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Glycosylation of the human erythrocyte glucose transporter is essential for glucose transport activity

Cited by 27 publications

References 23 publications

Selective Inhibition of Active Uptake of Sucrose into Plasma Membrane Vesicles by Polyclonal Sera Directed against a 42 Kilodalton Plasma Membrane Polypeptide

Selective Inhibition of Active Uptake of Sucrose into Plasma Membrane Vesicles by Polyclonal Sera Directed against a 42 Kilodalton Plasma Membrane Polypeptide

Structural and functional consequences of an N-glycosylation mutation (HEMPAS) affecting human erythrocyte membrane glycoproteins

Heterologous Expression and Functional Characterization of a Mouse Renal Organic Anion Transporter in Mammalian Cells

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