Synthesis of (/sup 3/H)phlorizin and its binding behavior to renal brush-border membranes
Journal Article
·
· Anal. Biochem.; (United States)
Tetra- and tribromophlorizin have been synthesized under mild brominating conditions. With catalytic debromination in the presence of hydrogen or tritium gas, bromine atoms in the derivatives were completely substituted by hydrogen or tritium. The product was identical to the native phlorizin and was chemically pure. Tritiated phlorizin with extremely high specific radioactivity (45 Ci/mmol) was obtained when hydrogen gas was replaced by tritium gas. While the brominated compounds showed little inhibition of sodium D-glucose co-transport by isolated renal brush-border membranes, (/sup 3/H)phlorizin had the same binding affinity to the brush-border membranes as native phlorizin and a K/sub i/ value of 1.2 ..mu..M for the sodium-dependent D-glucose transport. Binding studies performed using a flow-dialysis method resulted in 150 pmol of phlorizin-binding sites per milligram of membrane protein. This radioactive phlorizin can be a useful tool for determining D-glucose-(phlorizin) binding sites at a low phlorizin concentration in membranes, in nonvesicle forms such as collapsed membrane vesicles, and in purified protein fractions.
- Research Organization:
- Albert Einstein Coll. of Medicine, Bronx, NY
- OSTI ID:
- 5606151
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 129:2; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
BIOCHEMICAL REACTION KINETICS
BODY
BROMINATION
CARBOHYDRATES
CATALYSIS
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL BONDS
CHEMICAL REACTIONS
ELEMENTS
GLUCOSE
GLYCOSIDES
HALOGENATION
HEXOSES
HYDROGEN
ISOTOPE APPLICATIONS
KETONES
KIDNEYS
KINETICS
LABELLED COMPOUNDS
MEMBRANES
MONOSACCHARIDES
NONMETALS
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANS
PHLORIZIN
PROTEINS
REACTION KINETICS
RECEPTORS
SACCHARIDES
SYNTHESIS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TUBULES
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
BIOCHEMICAL REACTION KINETICS
BODY
BROMINATION
CARBOHYDRATES
CATALYSIS
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL BONDS
CHEMICAL REACTIONS
ELEMENTS
GLUCOSE
GLYCOSIDES
HALOGENATION
HEXOSES
HYDROGEN
ISOTOPE APPLICATIONS
KETONES
KIDNEYS
KINETICS
LABELLED COMPOUNDS
MEMBRANES
MONOSACCHARIDES
NONMETALS
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANS
PHLORIZIN
PROTEINS
REACTION KINETICS
RECEPTORS
SACCHARIDES
SYNTHESIS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TUBULES