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

From National Research Council Canada

Download
  1. (PDF, 1 MB)
DOIResolve DOI: https://doi.org/10.1099/mic.0.000221
AuthorSearch for: ; Search for: ; Search for: 1 ; Search for: 1 ; Search for: ; Search for: ; Search for:
Name affiliation
  1. National Research Council Canada. Human Health Therapeutics
FormatText
TypeArticle
Journal titleMicrobiology
ISSN1350-0872
1465-2080
Abstract
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. While the amount of FlaB2 was similar for cells grown below 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 nonarchaellated. Unexpectedly, a lower apparent molecular mass for FlaB2 was observed in western blots of cells grown at temperatures beyond 38°C suggestive of a truncation in the attached N-linked tetrasaccharide at higher growth temperatures. Mass spectrometry 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 wildtype glycan.
Publication date
LanguageEnglish
Peer reviewedYes
NPARC number21277136
Export citationExport as RIS
Report a correctionReport a correction
Record identifier01e07e5c-d779-42e3-9bc7-912a246b32b6
Record created2015-12-14
Record modified2019-03-06
Date modified:
Report a problem on this page