Abstract
Brush border membrane vesicles (BBMV) enriched in sucrase, maltase and alkaline phosphatase, and impoverished in Na+-K+-ATPase, were isolated from proximal and distal intestine of the gilthead sea bream (Sparus aurata) by a MgCl2 precipitation method. Vesicles were suitable for the study of the characteristics of D-glucose apical transport. Only one D-glucose carrier was found in vesicles from each intestinal segment. In both cases, the D-glucose transport system was sodium-dependent, phlorizin-sensitive, significantly inhibited by D-glucose, D-galactose, α-methyl-D-glucose, 3-O-methyl-D-glucose and 2-deoxy-D-glucose, and showed stereospecificity. Apparent affinity constants of D-glucose transport (Kt) were 0.24 ± 0.03 mM in proximal and 0.18 ± 0.03 mM in distal intestine. Maximal rate of influx (Jmax) was 47.3 ± 2.2 pmols. mg−1 protein for proximal and 27.3 ± 3.6 pmols. mg−1 protein for distal intestine. Specific phlorizin binding and relative abundance of an anti-SGLT1 reactive protein were significantly higher in proximal than in distal BBMV. These results suggest the presence of the same D-glucose transporter along the intestine, with a higher density in the proximal portion. This transporter is compatible with the sodium-dependent D-glucose carrier described for other fish and with the SGLT1 of higher vertebrates.
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Acknowledgments
This work has been supported by Spanish government (grant MAR97-0408-00-02 of CICYT) and Generalitat de Catalunya (grant 2001-SGR-00142 of CIRIT, and Centre de Referència en Aqüicultura). M. Sala-Rabanal was a recipient of a Formació de Personal Investigador grant from the Generalitat de Catalunya. The authors thank Dr. Carles Garriga and Anna Pérez-Bosque for their assistance in the performance of the Western blot and the immunohistochemichal analyses, respectively.
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Sala-Rabanal, M., Gallardo, M., Sánchez, J. et al. Na-dependent D-Glucose Transport by Intestinal Brush Border Membrane Vesicles from Gilthead Sea Bream (Sparus aurata). J Membrane Biol 201, 85–96 (2004). https://doi.org/10.1007/s00232-004-0710-y
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DOI: https://doi.org/10.1007/s00232-004-0710-y