Involvement of amino acid 36 in TM1 in voltage sensitivity in mouse Na+/glucose cotransporter SGLT1.
Human SGLT1 protein is an established sodium-glucose cotransporter. Despite widespread use of the mouse as a model organism, the mouse SGLT1 homologue has yet to be functionally characterized. Additionally, the crystal structure of a sugar transporter homologue, Vibrio SGLT, has recently been described, however, it offers limited information about the role of transmembrane segments outside of the core ligand binding domains. In particular, the amino acids in TM1 were not assigned in the structure. To examine the contribution of TM1 to the function of SGLT1, we have cloned and characterized the biophysical properties of SGLT1 from mouse, mSGLT1, and compared it to a clone containing an amino acid substitution in TM1, F36S. As predicted, both proteins formed functional Na+/sugar cotransporters, but F36S-mSGLT1 showed decreased rates of sugar uptake and decreased apparent affinities for both Na+ and sugar compared to mSGLT1. Analysis of pre-steady-state currents and comparison with the crystal structure of Vibrio SGLT provide plausible mechanisms to explain the differences in function of these two proteins. Our data suggest that amino acids in TM1, which are not involved in ligand binding and translocation pathways, significantly influence the functional properties of sodium-glucose carrier proteins.
Díez-Sampedro
Ana
A
Department of Physiology and Biophysics, University of Miami, School of Medicine, Miami, FL 33136, USA. adiezsampedro@med.miami.edu
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001693:Biological Transport, Active; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D015640:Ion Channel Gating; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary; D051381:Rats; D012964:Sodium / Q000378:metabolism*; D051269:Sodium-Glucose Transporter 1 / Q000235:genetics / Q000502:physiology*
D000596:Amino Acids; C495675:Slc5a1 protein, mouse; D051269:Sodium-Glucose Transporter 1; D012964:Sodium; D005947:Glucose