Properties of hexose-transport regulatory mutants isolated from L6 rat myoblasts.
A hexose-transport regulatory mutant (D1/S4) was isolated from L6 rat myoblasts on the basis of its resistance to detachment and cell lysis in the presence of antibody and complement. Growth studies indicated that D1/S4 cells had a slower doubling time (29 h) compared with the parental L6 cells (22 h). Furthermore, after 9 days growth, less than 1% cell fusion was observed with D1/S4 cells, whereas 95% cell fusion was observed with the L6 cells. When the parental L6 cells were starved of glucose or treated with anti-L6 antibody, a significant increase in the Vmax, of 2-deoxy-D-glucose (dGlc) and 3-O-methyl-D-glucose (MeGlc) transport was observed. Although glucose-grown D1/S4 cells possessed normal hexose-transport activity, the above treatments had no effect on dGlc and MeGlc transport in these cells. Electrophoresis and immunoblotting studies revealed that D1/S4 cells possessed decreased amounts of a 112 kDa plasma-membrane protein. It is conceivable that this protein may play a role in triggering the antibody- and glucose-starvation-mediated activation of hexose transport and in myogenic differentiation. Unlike D1/S4, mutant F72, a mutant defective in the high-affinity hexose-transport system, was found to possess normal amounts of the 112 kDa protein. Although glucose starvation has no effect on the hexose-transport activity in this mutant, its hexose transport activity can be increased by antibody treatment. These studies with mutants suggest the involvement of regulatory components in the activation of hexose transport.
D'Amore
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Department of Biochemistry, University of Western Ontario, London, Canada.
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D019325:3-O-Methylglucose; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001692:Biological Transport; D002455:Cell Division; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003837:Deoxy Sugars / Q000493:pharmacokinetics*; D003847:Deoxyglucose / Q000493:pharmacokinetics*; D005947:Glucose / Q000378:metabolism; D007122:Immunoelectrophoresis; D007700:Kinetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008757:Methylglucosides / Q000493:pharmacokinetics*; D008759:Methylglycosides / Q000493:pharmacokinetics*; D009132:Muscles / Q000276:immunology / Q000378:metabolism*; D009154:Mutation; D051381:Rats
D000906:Antibodies; D003837:Deoxy Sugars; D008565:Membrane Proteins; D008757:Methylglucosides; D008759:Methylglycosides; D019325:3-O-Methylglucose; D003847:Deoxyglucose; D005947:Glucose