Expression, Purification, and Assembly of Archaellum Subcomplexes of Sulfolobus acidocaldarius
The archaellum assembly machinery and its filament consist of seven proteins in the crenarchaeon Sulfolobus acidocaldarius. We have so far expressed, purified, and biochemically characterized four of these archaellum subunits, namely, FlaX, FlaH, FlaI, and FlaF. FlaX, FlaH, and FlaI tightly interact and form the archaellum motor complex important for archaellum assembly and rotation. We have previously shown that FlaH forms an inner ring within a very stable FlaX ring, and therefore FlaX is believed to provide the scaffold for the assembly of the archaellum motor complex. Here we describe how to express and purify FlaX and FlaH and how the double ring structure both form can be obtained.
Key wordsArchaea Archaellum Archaellum motor complex Type IV pili Motility Archaeal flagellum
P.C. received funding from an ERC starting grant (ARCHAELLUM, 311523), and P.T. was supported by funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No 686647 (MARA).
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