Figure 2: SepSecS A239T, T325S, and Y334C variants co-purify with the bacterial chaperone GroEL. | Scientific Reports

Figure 2: SepSecS A239T, T325S, and Y334C variants co-purify with the bacterial chaperone GroEL.

From: Structural basis for early-onset neurological disorders caused by mutations in human selenocysteine synthase

Figure 2

(a) Soluble fractions of the WT and pathogenic variants of SepSecS were analyzed by S200 SEC. WT SepSecS elutes as a tetramer in a symmetrical peak at ~70 min (−). The majority of Y334C (−−) elutes ~55 min, with a small fraction forming a tetramer. The entire sample of A239T (−•−) elutes at ~55 min, only a small fraction of T325S (•••) forms larger oligomeric species. Only traces recorded at 280 nm are shown for clarity. (b) The SDS-PAGE analysis shows that fractions eluting at ~70 min (lanes 1, 3, and 5) primarily contain SepSecS (MW ~60 kDa). The peaks eluting at ~55 min (lanes 2, 4, and 6) are impure and contain predominant bands at ~60 and 65 kDa. (c) Western-blot of SEC peaks reveals that the 60-kDa band is the His-tagged SepSecS, and that the 65-kDa band is the bacterial chaperone GroEL. The 70-min peak is SepSecS and the 55-min peak contains both GroEL and SepSecS.

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