Effects of growth conditions on archaellation and N-glycosylation in Methanococcus maripaludis

Microbiology (Reading). 2016 Feb;162(2):339-350. doi: 10.1099/mic.0.000221. Epub 2015 Dec 4.


In this study, the effects of growth conditions on archaellation in Methanococcus maripaludis were examined. Cells were grown in a variety of media, including complex, minimal and with formate as the electron donor, with different nitrogen sources, varied salinities and at a variety of growth temperatures. Of the conditions tested, Western blot results showed that major archaellin FlaB2 levels only varied detectably as a result of growth temperature. Whilst the amount of FlaB2 was similar for cells grown at < 35 °C, protein levels decreased at 38 °C and were barely detectable at 42 °C. Quantitative reverse transcription PCR experiments demonstrated that the flaB2 transcript levels were almost undetectable at 42 °C. Electron microscopy confirmed that the FlaB2 levels detected by Western blots corresponded to the state of archaellation, with cells grown at 42 °C being mostly non-archaellated. Unexpectedly, a lower apparent molecular mass for FlaB2 was observed in Western blots of cells grown at temperatures >38 °C, suggestive of a truncation in the attached N-linked tetrasaccharide at higher growth temperatures. MS analysis of archaella isolated from cells grown at 40 °C confirmed that FlaB2 was now decorated with a trisaccharide in which the third sugar was also lacking the attached threonine and acetamidino modifications found in the WT glycan.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Archaeal Proteins / genetics
  • Archaeal Proteins / metabolism*
  • Flagellin / genetics
  • Flagellin / metabolism*
  • Glycosylation
  • Hot Temperature
  • Methanococcus / growth & development*
  • Methanococcus / metabolism*
  • Microscopy, Electron, Transmission
  • Polysaccharides / metabolism*


  • Archaeal Proteins
  • FlaB2 protein, archaeal
  • Polysaccharides
  • Flagellin