Figure 3
From: Sarcolipin alters SERCA1a interdomain communication by impairing binding of both calcium and ATP
Effect of SLN during proteinase K treatment of recombinant purified SERCA1a (recS1a). (A) without SLN; (B) with palmitoylated SLN. Proteinase K proteolysis was performed in the presence of either 1 mM calcium (lanes 1–3) (cleavage at T242-E243) or EGTA and Thapsigargin (EG + Tg) (lanes 4–6) (cleavage at L119-K120) for 5 to 30 min before loading on SDS-PAGE as mentioned in the Methods section. Samples were also incubated for 30 min at 20 °C in absence of Proteinase K as standard, either in presence of 1 mM calcium (lanes 7–8) or EGTA and Tg (lanes 9–10), to assess the stability of the DDM-solubilized proteins. Thrombin, used for elution from the affinity chromatography resin, is present in the purified sample.
