SMIT2 mediates all myo-inositol uptake in apical membranes of rat small intestine.
This study presents the characterization of myo-inositol (MI) uptake in rat intestine as evaluated by use of purified membrane preparations. Three secondary active MI cotransporters have been identified; two are Na(+) coupled (SMIT1 and SMIT2) and one is H(+) coupled (HMIT). Through inhibition studies using selective substrates such as d-chiro-inositol (DCI, specific for SMIT2) and l-fucose (specific for SMIT1), we show that SMIT2 is exclusively responsible for apical MI transport in rat intestine; rabbit intestine appears to lack apical transport of MI. Other sugar transport systems known to be present in apical membranes, such as SGLT1 or GLUT5, lacked any significant contribution to MI uptake. Functional analysis of rat SMIT2 activity, via electrophysiological studies in Xenopus oocytes, demonstrated similarities to the activities of SMIT2 from other species (rabbit and human) displaying high affinities for MI (0.150 +/- 0.040 mM), DCI (0.31 +/- 0.06 mM), and phlorizin (Pz; 0.016 +/- 0.007 mM); low affinity for glucose (36 +/- 7 mM); and no affinity for l-fucose. Although these functional characteristics essentially confirmed those found in rat intestinal apical membranes, a unique discrepancy was seen between the two systems studied in that the affinity constant for glucose was approximately 40-fold lower in vesicles (K(i) = 0.94 +/- 0.35 mM) than in oocytes. Finally, the transport system responsible for the basolateral efflux transporter of glucose in intestine, GLUT2, did not mediate any significant radiolabeled MI uptake in oocytes, indicating that this transport system does not participate in the basolateral exit of MI from small intestine.
Aouameur
Rym
R
Groupe d'étude des protéines membranaires (GEPROM), Département de Physiologie, Université de Montréal, C.P. 6128, Succ. Centre-Ville, Montréal, Québec, Canada, H3C 3J7.
Da Cal
Sandra
S
Bissonnette
Pierre
P
Coady
Michael J
MJ
Lapointe
Jean-Yves
JY
D000818:Animals; D002478:Cells, Cultured; D007294:Inositol / Q000493:pharmacokinetics*; D007421:Intestine, Small / Q000378:metabolism* / Q000648:ultrastructure*; D008871:Microvilli / Q000378:metabolism; D051381:Rats; D051247:Sodium-Glucose Transport Proteins / Q000378:metabolism*
C526256:Slc5a11 protein, rat; D051247:Sodium-Glucose Transport Proteins; D007294:Inositol