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Preparation and characterization of basolateral membrane vesicles from pig and human colonocytes: the mechanism of glucose transport.
- Source :
-
The Biochemical journal [Biochem J] 1993 Sep 01; Vol. 294 ( Pt 2), pp. 529-34. - Publication Year :
- 1993
-
Abstract
- Membrane vesicles were isolated from the basolateral domains of pig and normal human colonocytes. The activity of the ouabain-sensitive K(+)-activated phosphatase, the basolateral membrane marker, was enriched 13-fold in these membrane vesicles over the original homogenate. The membranes displayed cross-reactions with antibodies to the (Na+/K+)ATPase and the RLA class I major histocompatibility antigen, both known indicators of the basolateral membrane. There was negligible contamination by other organelles and the luminal membrane, as revealed by marker-enzyme analysis and Western blotting, using an antibody to villin. The vesicles transported D-glucose in a cytochalasin B-inhibitable Na(+)-independent manner, with a Km of 28.1 +/- 0.8 mM and Vmax. of 3.1 +/- 0.4 nmol/s per mg of protein. The transport was inhibited by 2-deoxy-D-glucose and 3-O-methyl-D-glucose, but not by L-glucose or methyl-alpha-D-glucose. Probing the colonocyte basolateral membranes with an antibody against the C-terminus of the human liver GLUT 2 produced a cross-reaction at 52 kDa. These properties indicate the presence of a GLUT 2 isoform on the basolateral membranes of human and pig colonocytes.
- Subjects :
- 3-O-Methylglucose
Alkaline Phosphatase analysis
Animals
Biological Transport drug effects
Cell Fractionation
Cell Membrane chemistry
Cell Membrane ultrastructure
Cytochalasin B pharmacology
Deoxyglucose pharmacology
Glucose Transporter Type 2
Histocompatibility Antigens Class I analysis
Humans
Methylglucosides pharmacology
Monosaccharide Transport Proteins analysis
Ouabain pharmacology
Potassium pharmacology
Sodium pharmacology
Sodium-Potassium-Exchanging ATPase analysis
Swine
Cell Membrane metabolism
Colon ultrastructure
Glucose metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0264-6021
- Volume :
- 294 ( Pt 2)
- Database :
- MEDLINE
- Journal :
- The Biochemical journal
- Publication Type :
- Academic Journal
- Accession number :
- 8396917
- Full Text :
- https://doi.org/10.1042/bj2940529