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KMID : 1235920100020010021
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Medical Journal of Catholoc University of Daegu
2010 Volume.2 No. 1 p.21 ~ p.26
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Hexose Inhibition of 2-deoxy-D-Glucose Transport of the Endogenous Glucose Transport Systems in High Five Cells
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Lee Chong-Kee
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Abstract
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Background: Unlike mammalian transporters, little is known about the nature of the sugar transporter (s) in insect cells. In order to examine the transport characteristics of the insect cell transporter (s), the affinities of the insect cell transporter for different hexoses were assessed by examining the ability of these sugars to inhibit 2-deoxy-D-glucose (2dGlc) transport in High Five cells, which are used as a host cell for the baculovirus replication.
Materials and Methods: The hexose transport characteristics of High Five cells were investigated using 2-[1,2-3H]-2dGlc. Since the same substrate (2dGlc) was used for all experiments of this type, then the apparent Ki values for each sugar should provide a measure of their relative affinities.
Results: The Ki ‘s for reversible inhibitors were determined from plots of uptake versus inhibitor concentration.
Transport was effectively inhibited by D-glucose, but not L-glucose. Of the hexoses tested, L-glucose had the least effect on 2dGlc transport in the insect cells.
Conclusion: Like the human erythrotype type glucose transport system, D-glucose had a greater affinity for the insect cell transporter than L-glucose, indicating that the transport is stereoselective. In addition, epimerization at the C-4 position of D-glucose (D-galactose) resulted in a dramatic decrease in affinity of the hexose for the insect cell transporter. It is therefore concluded that the insect cells were appear to contain an endogenous sugar transport activity that in several aspects resembles the human erythrocyte type glucose transporter, although the human and insect transporters do differ in their affinity for cytochalasin B.
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KEYWORD
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Insect cells, Hexose transport, Transport assay
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