Contribution of cysteines C59 and C61 in the first transmembrane helix (TM1) to the stabilization of the ASIC1a complex by BMOE. Miguel Xavier van Bemmelen Delphine Huser Ivan Gautschi Laurent Schild 10.1371/journal.pone.0135191.g004 https://plos.figshare.com/articles/figure/_Contribution_of_cysteines_C59_and_C61_in_the_first_transmembrane_helix_TM1_to_the_stabilization_of_the_ASIC1a_complex_by_BMOE_/1506177 <p>A: ASIC1a oligomeric states, resolved by SDS-PAGE under reducing conditions and detected by anti-His-tag western blotting (same experimental procedure as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0135191#pone.0135191.g003" target="_blank">Fig 3A</a>), of the His<sub>8</sub>-tagged forms of ASIC1a-ΔC<sub>Ct</sub> as control (Ctrl), and the cysteine substitution mutant G433C-ΔC<sub>Ct</sub> (433), G433C-C59V-ΔC<sub>Ct</sub> (433–59) or G433C-C59V-C61S-ΔC<sub>Ct</sub> (433-59-61). Numbers I to IV have the same meaning as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0135191#pone.0135191.g003" target="_blank">Fig 3</a>. B: Relative intensities (mean ±SEM, n = 4) of each of the 4 bands (I to IV) for ASIC1a identified on SDS-PAGE from cells expressing ASIC1a-ΔC<sub>Ct</sub>, (C), G433C-ΔC<sub>Ct</sub> (433) G433C-C59V-ΔC<sub>Ct</sub> (433–59) and G433C-C59V-C61S-ΔC<sub>Ct</sub> (433-59-61). The average molecular weight estimated for the four bands I, II, III, IV are respectively 80±10, 160±6, 230±9 and 300±19 kDa (Mw±SD, n = 4) for the G433C-ΔC<sub>Ct</sub>, G433C-C59V-ΔC<sub>Ct</sub>, and G433C-C59V-C61S-ΔC<sub>Ct</sub> constructs. * denotes p<0.01</p> 2015-08-07 04:03:38 extracellular pore vestibule oligomeric states ASIC 1a complexes Xenopus laevis oocytes sulfhydryl crosslinker BMOE Homotetrameric Assembly State ASIC 1a monomers crosslinking conditions ASIC 1a ASIC 1a oligomers ASIC 1a subunit cell surface ASIC 1a channel tetrameric concatemeric cDNA cho ASIC 1a homotetramer ASIC 1 complexes