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