title: Compound missense mutations in the sodium/D-glucose cotransporter result in trafficking defects.
abstract: OBJECTIVE
  Defects in the Na+-dependent glucose transporter (SGLT1) are associated with the disorder glucose-galactose malabsorption, characterized by severe diarrhea. This study focused on a unique proband with glucose-galactose malabsorption who was investigated 30 years ago, and the aims of the study were to identify mutations in the SGLT1 gene and to determine the defect in sugar transport.
  
  
  METHODS
  Mutations were identified by sequencing, and each mutant protein was then studied using a Xenopus oocyte heterologous expression system. Analysis included Western, freeze fracture, radiotracer uptake, and electrophysiological assays.
  
  
  RESULTS
  Two heterozygous missense mutations (Cys355Ser and Leu147Arg) were identified that entirely eliminated Na+/sugar cotransport activity. Western blot analysis showed that the levels of both mutant proteins in the oocyte were comparable to wild-type SGLT1, but no complex glycosylation was detected. No SGLT1 charge movements were observed with the mutant proteins, and freeze fracture data showed that neither mutant protein reached the plasma membrane.
  
  
  CONCLUSIONS
  The Cys355Ser and Leu147Arg mutations eliminate the Na+/sugar cotransport by blocking the transfer of SGLT1 protein from the endoplasmic reticulum to the plasma membrane. This is consistent with earlier studies on phlorizin binding to the brush border membrane of duodenal biopsy specimens from this patient.
authors.affiliation: Department of Pediatrics, UCLA School of Medicine, Los Angeles, California 90095-1751, USA. martin@physiology.medsch.ucla.edu
authors.forename: M G
authors.initials: MG
authors.lastname: Martín
authors.forename: M P
authors.initials: MP
authors.lastname: Lostao
authors.forename: E
authors.initials: E
authors.lastname: Turk
authors.forename: J
authors.initials: J
authors.lastname: Lam
authors.forename: M
authors.initials: M
authors.lastname: Kreman
authors.forename: E M
authors.initials: EM
authors.lastname: Wright
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D004594:Electrophysiology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005260:Female; D006801:Humans; D007231:Infant, Newborn; D008286:Malabsorption Syndromes / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D008757:Methylglucosides / Q000493:pharmacokinetics; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D009154:Mutation; D009865:Oocytes / Q000378:metabolism / Q000648:ultrastructure; D010375:Pedigree; D051269:Sodium-Glucose Transporter 1; D014982:Xenopus laevis
chemical_list: D008562:Membrane Glycoproteins; D008757:Methylglucosides; D009004:Monosaccharide Transport Proteins; C495674:SLC5A1 protein, human; D051269:Sodium-Glucose Transporter 1; C027020:methylglucoside