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

  • Paushali Chaudhury
  • Patrick Tripp
  • Sonja-Verena Albers
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Archaea Archaellum Archaellum motor complex Type IV pili Motility Archaeal flagellum 

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Paushali Chaudhury
    • 1
  • Patrick Tripp
    • 1
    • 2
  • Sonja-Verena Albers
    • 1
  1. 1.Molecular Biology of Archaea, Institute of Biology IIUniversity of FreiburgFreiburgGermany
  2. 2.Spemann Graduate School of Biology and Medicine (SGBM)University of FreiburgFreiburgGermany

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