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초록· 키워드
The baculovirus expression system is a powerful method for producing large amounts of the human erythrocyte-type glucose transport protein, heterologously. Characterization of the expressed protein is expected to show its ability to transport sugars directly. To achieve this, it is a prerequisite to how the properties of the endogenous sugar transport system in Spodoptera frugiperda Clone 21 (Sf21) cells, which are commonly employed as a host permissive cell line to support the baculovirus replication, The Sf21 cells can grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transport system. However, unlike the human glucose transport protein that has a broad substrate and inhibitor specificity, very little is hewn about the nature of the endogenous sugar transport system in Sf21 cells. In order to characterize further the inhibitor recognition properties of the Sf21 cell transporter, the ability of phloretin, cytochalasin B and D-fructose to inhibit 2-deoxy-D-glucose (2dGlc) transport was examined by measuring inhibition constants (K<sub>i</sub>). The K<sub>i</sub>'s for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. The 2dGlc transport in the Sf21 cells was very potently inhibited by phloretin, the aglucone of phlorizin with a K<sub>i</sub> similar to the value of about 2 μM reported for inhibition of glucose transport in human erythrocytes. However, the Sf21 cell transport system was found to differ from the human transport protein in being much less sensitive to inhibition by cytochalasin B (apparent K<sub>i</sub> of approximately 10 μM). In contrast, It is reported that the inhibitor binds the human erythrocyte counterpart with a K<sub>d</sub> of approximately 0.12 μM. Interestingly, the Sf21 glucose transport system also appeared to have high affinity (3r D-fructose with a K<sub>i</sub> of approximately 5 mM, contrasting the reported K<sub>m</sub> of the human erythrocyte transport protein for the ketose of 1.5 M.
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UCI(KEPA) : I410-ECN-0101-2009-510-017357015
