Analysis of low temperature tolerance of membrane transporters and development of pan-transporter inhibitors

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Low temperature tolerance of membrane transporters, a characteristic showing transport activity even at ice-cold temperature, was examined in human erythrocytes and cultured cancer cells. A facilitative glucose transporter GLUT1 and a facilitative nucleoside transporter ENT1 showed marked low temperature tolerance in erythrocyte membrane vesicles. A secondary active monocarboxylate transporter MCT1 showed low temperature tolerance in erythrocytes. In addition, GLUT1 in culture cells such as A549 and HepG2 also showed low temperature tolerance.In order to evaluate the function of these transporters properly, I attempted to develop novel pan-transporter inhibitors (stop and washing solutions for transport study). Among various reagents tested, solutions containing urea plus formaldehyde and 2-mercaptoethanol plus glutaraldehyde were found to be effective pan-transporter inhibitors in both membrane vesicle and culture cell studies.