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Expression, Purification, and Assembly of Archaellum Subcomplexes of Sulfolobus acidocaldarius

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Protein Complex Assembly

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1764))

Abstract

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.

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Acknowledgment

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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Correspondence to Sonja-Verena Albers .

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Chaudhury, P., Tripp, P., Albers, SV. (2018). Expression, Purification, and Assembly of Archaellum Subcomplexes of Sulfolobus acidocaldarius . In: Marsh, J. (eds) Protein Complex Assembly. Methods in Molecular Biology, vol 1764. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7759-8_19

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  • DOI: https://doi.org/10.1007/978-1-4939-7759-8_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7758-1

  • Online ISBN: 978-1-4939-7759-8

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