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. 2006 May;127(5):467–480. doi: 10.1085/jgp.200509467

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Copyright © 2006, The Rockefeller University Press

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Figure 8. — Protection of pore-lining cysteine residues by putrescine. Patches expressing (A) Kir6.2[N160D][C166S][L157C], (B) Kir6.2[N160D][C166S][L164C], or (C) Kir6.2[N160D][C166S][L169C] were blocked at +50 mV in 1 mM putrescine, exposed to 100 μM MTSEA for a variable interval (while continuously exposed to putrescine), and repolarized to −50 mV to determine the extent of MTSEA modification. Sample traces for each construct are presented in the lefthand panels, along with the unprotected MTSEA modification rates for comparison. Compiled data from multiple patches are presented in the righthand panels, and fit with a single exponential curve. Unprotected modification time courses are indicated by the dashed blue lines. At residue 157C, putrescine slowed the time constant of modification to 15.8 ± 1.3 s, from an unprotected time constant of 4.2 ± 0.7 s. At residue 164C in the presence of putrescine, the modification time constant was 7.1 ± 0.5 s, and the unprotected time constant was 3.9 ± 0.5 s. At residue 169C, putrescine slowed the modification time constant to 3.6 ± 0.3 s, from the unprotected time constant of 2.4 ± 0.3 s.