Reconstruction of central carbon metabolism in Sulfolobus solfataricus using a two-dimensional gel electrophoresis map, stable isotope labelling and DNA microarray analysis

B.P.L. Snijders, J. Walther, S. Peter, I. Kinnman, M.J.G. de Vos, H.J.G. van de Werken, S.J.J. Brouns, J. van der Oost, P.C. Wright

Research output: Contribution to journalArticleAcademicpeer-review

52 Citations (Scopus)

Abstract

In the last decade, an increasing number of sequenced archaeal genomes have become available, opening up the possibility for functional genomic analyses. Here, we reconstructed the central carbon metabolism in the hyperthermophilic crenarchaeon Sulfolobus solfataricus (glycolysis, gluconeogenesis and tricarboxylic acid cycle) on the basis of genomic, proteomic, transcriptomic and biochemical data. A 2-DE reference map of S. solfataricus grown on glucose, consisting of 325 unique ORFs in 255 protein spots, was created to facilitate this study. The map was then used for a differential expression study based on (15)N metabolic labelling (yeast extract + tryptone-grown cells (YT) vs. glucose-grown cells (G)). In addition, the expression ratio of the genes involved in carbon metabolism was studied using DNA microarrays. Surprisingly, only 3 and 14% of the genes and proteins, respectively, involved in central carbon metabolism showed a greater than two-fold change in expression level. All results are discussed in the light of the current understanding of central carbon metabolism in S. solfataricus and will help to obtain a system-wide understanding of this organism.
Original languageEnglish
Pages (from-to)1518-1529
JournalProteomics
Volume6
Issue number5
DOIs
Publication statusPublished - 2006

Keywords

  • entner-doudoroff pathway
  • citric-acid cycle
  • hyperthermophilic archaea
  • archaebacterium sulfolobus
  • citrate synthase
  • phosphoenolpyruvate carboxylase
  • thermoacidophilic crenarchaeon
  • thermoplasma-acidophilum
  • thermoproteus-tenax
  • glycolytic pathways

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