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. Author manuscript; available in PMC: 2010 Oct 15.
Published in final edited form as: Biochim Biophys Acta. 2007 Nov 29;1778(2):454–465. doi: 10.1016/j.bbamem.2007.11.010

Fig. 5. ATP-dependent LTC4 uptake by membrane vesicles prepared from wild-type and mutant MRP1s.

Fig. 5

A. Relative amounts of MRP1 proteins in membrane vesicles containing wild-type or mutant MRP1s. The membrane vesicles containing mutant MRP1s were prepared from cells grown at 27 °C so that majorities of the mutant MRP1s are complex-glycosylated mature proteins. The amounts of membrane vesicle proteins loaded in the gel are indicated on the top. MRP1 on the left indicates the 190 kDa complex-glycosylated mature MRP1 proteins that were detected in western blot by employing the monoclonal antibody 42.4 [20]. The intensities of the 190 kDa bands were determined by a scanning densitometer. The mean ratios (n = 3), considering the amount of wild type MRP1 as 1, of the mutant proteins are listed at the bottom of the gel. B. Relative rate of ATP-dependent LTC4 transport activity. The assays were carried out in a 30 μl solution containing 3 μg of membrane vesicles (the amount of MRP1 protein determined in panel A was adjusted to a similar amount by adding varying amount of membrane vesicles prepared from parental BHK cells) and 4 mM AMP (used as a control) or 4 mM ATP at 37 °C for 4 minutes. The amount of LTC4 bound to the membrane vesicles in the presence of 4 mM AMP was subtracted from the corresponding samples in the presence of 4 mM ATP. The data are means ± S.D. of four triplicate-determinations.