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Transport and subcellular distribution characteristics of human erythrocyte glucose transporter fused in adipocytes

Thesis/Dissertation ·
OSTI ID:5545227
Purified human erythrocyte glucose transporters (HEGT) were incorporated into the rat epididymal adipocytes by polyethylene glycol (PEG)-induced fusion. The incorporation of HEGT was found to be dependent on both the molecular weight and the concentration of PEG. Optimal incorporation of HEGT occurred when 10% PEG 8000 was used. This incorporation was found to be proportional to the increasing amounts of HEGT used. Morphology tests showed very little surface adsorption of HEGT on adipocytes after fusion. Transport activity of fused HEGT was studied by measuring equilibrium exchange of 3-O-methylglucose. In adipocytes employed this fusion protocols, the transport rate increased significantly compared with cells treated under non-fusion conditions. This fold increase was directly proportional to the amount of HEGT recovered in the plasma membrane (PM). Furthermore, calculated turnover number of HEGT in fused adipocytes was as high as that of the native adipocyte glucose transporter (AGT). Subcellular distribution of HEGT in fused adipocytes was assessed using ({sup 3}H)cytochalasin B-labeled HEGT vesicles. HEGT was distributed in each subcellular organelle at specific ratios. This relative distribution was almost constant regardless of the amount of HEGT used. The distributions of lipid-labeled HEGT, fluid-phase endocytosed HEGT and free HEGT mixed to adipocyte homogenate were shown to be completely different from that of protein-labeled HEGT fusion.
Research Organization:
State Univ. of New York, Buffalo, NY (United States)
OSTI ID:
5545227
Country of Publication:
United States
Language:
English

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Related Subjects

550501* -- Metabolism-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
ANIMAL CELLS
ANIMALS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOHYDRATES
CELL CONSTITUENTS
CELL MEMBRANES
CONNECTIVE TISSUE CELLS
DISTRIBUTION
ERYTHROCYTES
FAT CELLS
GLUCOSE
HEXOSES
HYDROGEN COMPOUNDS
ISOTOPE APPLICATIONS
MAMMALS
MAN
MATERIALS
MEMBRANE TRANSPORT
MEMBRANES
METABOLISM
MOLECULAR WEIGHT
MONOSACCHARIDES
ORGANIC COMPOUNDS
PRIMATES
PROTEINS
RATS
RODENTS
SACCHARIDES
SOMATIC CELLS
SUBCELLULAR DISTRIBUTION
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES