Na-dependent D-glucose transport by intestinal brush border membrane vesicles from gilthead sea bream (Sparus aurata)

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, alpha-methyl-D-glucose, 3-O-methyl-D-glucose and 2-deoxy-D-glucose, and showed stereo specificity. Apparent affinity constants of D-glucose transport (K-t) 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.