The acetate-uptake ability of MBA4 was inhibited by propionate but not by butyrate. This is consistent with the acetate permease ActP of E. coli [17]. The failure of butyrate and valerate

to act as a competing solute suggested that a molecule with more than three carbons would be less effective for the acetate-uptake system. In summary, Trametinib datasheet MCA, MBA, 2MCPA, and butyrate could inhibit MCA- but not acetate- uptake of MBA4. A visual inspection of the structural models of these molecules (Figure 7) suggests that they are generally larger than acetate. Similarly, MCA, MBA, and propionate have a stronger inhibitory effect on MCA uptake than 2MCPA and butyrate. The failure of valerate to act as a competing solute further strengthens the DNA/RNA Synthesis inhibitor notion that size is a determining factor. By means of comparing the structures of the competing solutes it may be possible to estimate the range of substrates recognized by various transport systems and provide valuable information on the functional property of the transporters. Figure 7 Structural models mTOR inhibitor of various competing solutes. The values of atomic radii, the skeletal formula and the space-filling models of acetic acid, MCA, MBA, propionic acid, 2MCPA, butyric acid, and valeric acid were obtained from ACD/ChemSketch (Advanced Chemistry Development, Inc.). The solutes were assumed to be in disassociated

forms in PBS buffer (pH 7.4) used in this study. The inducers for the acetate-uptake system are acetate, MCA, MBA, propionate, and 2MCPA, but only acetate and propionate are substrates. Similarly, the inducers for the MCA uptake system are MCA, MBA and to a lesser extend 2MCPA, while the substrates include the inducers, acetate and propionate. The inducer and the substrate are not necessarily the same. Although the acetate- and the MCA- transport systems have different induction patterns and substrate

specificities, they do share certain similarity. The activities of both systems were abolished by CCCP, suggesting transmembrane electrochemical potential as a driving force. As CCCP could not discriminate between proton- and sodium-coupled symport, it was unclear which was/were involved in the transports. Previous studies of bacterial acetate-transport systems failed to give a uniformed conclusion. Although ActP of E. coli was assigned to the sodium:solute Carbachol symporter family, no dependency on sodium was demonstrated [17]. While electrochemical proton potential was confirmed to be a driving force for MctC of Corynebacterium glutamicum [18], acetate uptake in Accumulibacter spp. was believed to be driven by proton motive force, and in Defluviicoccus spp. it was suggested to be powered by proton or sodium gradient or both [23]. An increased uptake of acetate for a change of pH from 8 to 4 affirmed the involvement of proton in acetate transport in MBA4. However, the involvement of sodium could not be ruled out and further confirmation is required.