Expression and molecular characterization of rat renal D-mannose transport in Xenopus oocytes

J Membr Biol. 2000 Nov 15;178(2):127-35. doi: 10.1007/s002320010020.

Abstract

Renal reabsorption appears to play a major role in d-mannose homeostasis. Here we show that in rat kidney, the transport of d-mannose by brush border membrane vesicles from tubular epithelial cells involves an uphill and rheogenic Na-dependent system, which is fully inhibited by d-mannose itself, incompletely inhibited by d-glucose, d-fructose, phloridzin, and phloretin, and noninhibited by l-mannose or disaccharides. In addition, this system exhibits both low capacity (112.9+/-15.6 pmol/mg/second) and high affinity (0.18+/-0.04 mm), with a 2:1 stoichiometry for the Na:d-mannose interaction, and low affinity for sodium (16.6+/-3.67 mm). We also show expression of d-mannose transport by Xenopus laevis oocytes injected with rat renal polyA(+) RNA. Kinetic analysis of the expressed transport was performed after RNA enrichment by fractionation through a sucrose density gradient and was shown to be identical to that measured in membrane vesicles. The RNA species encoding the expressed transport has a small mean size, 1 kb approximately, and shows no homology with the SGLT family of Na-dependent d-glucose transporters, as shown by low stringent RT-PCR and northern analysis. The expressed transport is specific for d-mannose, since in spite of a significant inhibition by d-glucose and d-fructose, neither of these two substrates was transported above the level of the water-injected oocytes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Transport, Active / drug effects
  • Cell Fractionation
  • Fructose / pharmacology
  • Glucose / pharmacology
  • Kidney Cortex / metabolism*
  • Kinetics
  • Mannose / metabolism*
  • Membrane Glycoproteins / genetics
  • Microvilli / metabolism
  • Monosaccharide Transport Proteins / genetics
  • Oocytes / drug effects
  • Oocytes / metabolism
  • Phloretin / pharmacology
  • Phlorhizin / pharmacology
  • RNA, Messenger / isolation & purification
  • RNA, Messenger / pharmacology
  • Rats
  • Rats, Wistar
  • Sodium-Glucose Transporter 1
  • Sucrose / chemistry
  • Transport Vesicles / metabolism
  • Xenopus / genetics
  • Xenopus / metabolism*

Substances

  • Membrane Glycoproteins
  • Monosaccharide Transport Proteins
  • RNA, Messenger
  • Sodium-Glucose Transporter 1
  • Fructose
  • Sucrose
  • Phlorhizin
  • Glucose
  • Mannose
  • Phloretin