Summary
Ultimately the distribution of control among the various factors in the cell depends on the kinetic properties of all the components in the cell. These properties may again differ between the isolated state and the intact system. Consequently it is important to be able to study the kinetic properties both in well-defined states and in the intact cell.
We review our studies on the function of P-glycoprotein as measured in intact cells by monitoring daunorubicin fluorescence in a flow-through system. To interpret the experimental findings, a mathematical model was set up. Multidrug resistance is not only controlled by the drug efflux pump. Part of the control resides in the passive permeability of the plasma membrane for the drug.
The control distribution between pump and the passive permeability should depend on how the pumping rate of P-glycoprotein depends on the intracellular daunorubicin concentration. Interestingly, the latter dependence was more than linear. This co-operativity in terms of daunorubicin concentration was confirmed when studying active drug pumping into plasma membrane vesicles loaded with DNA, by following fluorescence quenching of the transported daunorubicin.
The intricate organisation of cellular processes brings with it special kinetic and control properties. Here we discuss this for the case of ‘vacuum cleaning’multidrug pumps and for the bacterial phosphotransferase system (PTS), which combines transport with regulation. The distinction between ‘vacuum cleaning’ and ‘non vacuum cleaning’ multidrug pumps may be less strict than anticipated and we define a “Vacuum Cleaner Index”which may serve to quantify the extent to which a drug pump behaves as a “vacuum cleaner”. Using the PTS as a model system for channeling and signal transduction, we show experimentally that the sum of the flux control.coefficients of the PTS enzymes may come close 2.
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Westerhoff, H.V. et al. (1996). Direct Transfer of Control and Multidrug Resistance. In: Kohen, E., Hirschberg, J.G. (eds) Analytical Use of Fluorescent Probes in Oncology. NATO ASI Series, vol 286. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5845-3_29
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DOI: https://doi.org/10.1007/978-1-4615-5845-3_29
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