Fructose transport and GLUT-5 protein in human sarcolemmal vesicles.

Abstract

Sarcolemmal vesicles were produced from human skeletal muscle biopsy material obtained at rest and immediately after maximal dynamic exercise (100% maximal O2 uptake) for analysis of fructose transport and hexose transporter (GLUT-5) protein concentration. Human sarcolemmal vesicles displayed a time-dependent uptake of D-fructose that displayed saturable Michaelis-Menten type kinetics (maximal transport 477 +/- 37 pmol.min-1.mg protein-1; half-maximal concentration constant 8.3 +/- 1.2 mM). At a hexose concentration of 5 mM, vesicle transport rate was eight times faster for glucose than for fructose. Preincubation of human muscle vesicles with 35 microM cytochalasin B before the uptake assay resulted in > 95% inhibition in D-glucose uptake, whereas transport of D-fructose was unaffected. Sarcolemmal vesicles prepared from exercised human muscle showed a significant increase (49%) in vesicle GLUT-4 content (P < 0.03, n = 10), which accounts for the increase in vesicle glucose transport that we have recently reported [S. Kristiansen, M. Hargreaves, and E.A. Richter. Am. J. Physiol. 270 (Endocrinol. Metab. 33): E197-E201, 1996]. In contrast, exercise did not increase the vesicle GLUT-5 protein content or induce changes in vesicle fructose transport activity. In conclusion, we propose that fructose transport into human skeletal muscle occurs via a mechanism distinct from that utilized by glucose on the basis of differences in sensitivity to cytochalasin B and responsiveness to exercise. Furthermore, our findings signify that uptake of fructose in human skeletal muscle is mediated by the GLUT-5 transporter.