Interaction of facilitative glucose transporter with glucokinase and its modulation by ADP and glucose-6-phosphate.
Bacterial glucokinase (GK) binds to purified, human erythrocyte glucose transporter (GT) reconstituted in vesicles. The binding is largely abolished if GT is predigested with trypsin, indicating that GK binds to the cytoplasmic domain of GT. The binding is a saturable function of GK concentration showing two distinct affinities with apparent KD of 0.33 and 5.1 microM. The binding is stimulated by an increasing concentration of ADP with the 50% maximal effect at 5 mM. Glucose-6-phosphate (G6P) also stimulates the binding with a distinct optimum at 25 mM. The binding is stimulated only slightly by ATP. D-glucose has no affect on the binding. KCl enhances the binding with the maximal effect at physiological intracellular concentrations. The binding is sensitive to changes in pH with an optimum at pH 4. The binding causes no detectable functional change in GT. However, the enzymatic activity of GK measured at nanomolar concentrations of GK is significantly greater in the presence of GT vesicles than in its absence or in the presence of protein-free vesicles, indicating that GK interacts with GT at this low concentration range with an apparent KD of 10 mM. Although its physiological significance is not known, the GK-GT interaction in vitro described here suggests that these two proteins may also interact in the cell and regulate carbohydrate metabolism.
Lachaal
M
M
Veterans Administration Medical Center, Buffalo, New York.
Jung
C Y
CY
D000244:Adenosine Diphosphate / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D005941:Glucokinase / Q000378:metabolism* / Q000502:physiology; D005947:Glucose / Q000378:metabolism; D051272:Glucose Transporter Type 1; D019298:Glucose-6-Phosphate; D005958:Glucosephosphates / Q000494:pharmacology*; D006593:Hexokinase / Q000502:physiology; D006706:Homeostasis / Q000502:physiology; D006801:Humans; D009004:Monosaccharide Transport Proteins / Q000032:analysis / Q000378:metabolism* / Q000502:physiology; D011189:Potassium Chloride / Q000494:pharmacology; D011485:Protein Binding; D014357:Trypsin / Q000494:pharmacology
D051272:Glucose Transporter Type 1; D005958:Glucosephosphates; D009004:Monosaccharide Transport Proteins; C495677:SLC2A1 protein, human; D019298:Glucose-6-Phosphate; D000244:Adenosine Diphosphate; D011189:Potassium Chloride; D006593:Hexokinase; D005941:Glucokinase; D014357:Trypsin; D005947:Glucose