Effects of Serum on Membrane Transport

When monolayers of freshly obtained rabbit lung macrophages are exposed to the nucleoside analogue, showdomycin (sho), adenosine transport, measured over a 45 s interval, is irreversibly inhibited. Low doses of the drug or short periods of exposure, however, do not result in decreased transport, while higher concentrations or longer exposures result in exponential decline. The initial lag is not due to a long reaction time of sho with the transport carrier or to nonspecific sites absorbing the drug. Previously it was shown that preincubation of monolayers with normal rabbit serum (NRS) results in increased adenosine transport. When monolayers are first exposed to sho so as to inhibit transport to varying degrees and then incubated with NRS, transport is increased over the inhibited level. Several experiments make it unlikely that serum removes the drug from the cell surface in a nonspecific fashion. Moreover, serum given before, during, or after sho alters the dose response curve so that no shoulder is seen. One way to explain these results makes use of target theory: the adenosine transport system could be comprised mainly of "coupled" or "clustered" sites of which only one is active at any time as well as "hidden" sites which are inactive. When a site in a group is irreversibly inactivated by sho, another in the group becomes activated. Serum might activate or uncouple all sites and also cause the appearance of hidden sites, which previously neither transported nor bound sho.