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. 2002 Nov;120(5):693–705. doi: 10.1085/jgp.20028645

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Copyright © 2002, The Rockefeller University Press

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Figure 2. — Effect of MTS reagents on electrogenic response of C-C mutant. (A) Representative current recordings from oocytes that expressed the single mutants A203C, S460C, and WT NaPi-IIa, before and after incubation in 100 μM MTSEA, MTSES, or MTSET (data shown only for MTSEA as MTSET and MTSES gave similar records). Traces show responses of oocytes to 1 mM P_i (filled bar) and 3 mM PFA (open bar) applied for the period indicated, before (−, left traces) and after (+, right traces) incubation. The continuous line indicates the holding current level reached during the initial PFA application. Dashed line represents holding current (I _hold) in ND100 solution. Note that for each cell, the recording baseline levels were not adjusted before and after MTSEA exposure. Note the quantification of transport modes as indicated: substrate-induced currents (I _PFA, I _Pi) are measured relative to I _hold. Uniport or leak activity is then given by −I _PFA and the cotransport activity is given by I _Pi − I _PFA. We assume that these substrate concentrations are sufficient to inhibit fully the leak current and that both transport modes are mutually exclusive. Bars: vertical, 50 nA; horizontal, 20 s. (B) Recordings from oocytes expressing the double mutant A203C-S460C (C-C) before (−, left traces) and after (+, right traces) incubation for 3 min with 100 μM MTSEA, MTSES, or MTSES. Note that for the double mutant, the leak current increases only in the presence of MTSEA and MTSES. Bars: vertical, 50 nA; horizontal, 20 s. (C) Pooled data for the leak for oocytes expressing WT, A203C, S460C, and C-C mutant after incubation with MTSEA (white bars), MTSES (black bars), or MTSET (gray bars). Each data point represents mean of five oocytes, normalized to the value before MTS exposure.