Purification and biochemical characterization of the ATP synthase from Heliobacterium modesticaldum

Jay How Yang, Iosifina Sarrou, Jose M. Martin-Garcia, Shangji Zhang, Kevin Redding, Petra Fromme

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Heliobacterium modesticaldum is an anaerobic photosynthetic bacterium that grows optimally at pH 6-7 and 52°C and is the only phototrophic member of the Firmicutes phylum family (gram-positive bacteria with low GC content). The ATP synthase of H. modesticaldum was isolated and characterized at the biochemical and biophysical levels. The isolated holoenzyme exhibited the subunit patterns of F-type ATP synthases containing a 5-subunit hydrophilic F<inf>1</inf> subcomplex and a 3-subunit hydrophobic F<inf>0</inf> subcomplex. ATP hydrolysis by the isolated HF<inf>1</inf>F<inf>0</inf> ATP synthase was successfully detected after pretreatment with different detergents by an in-gel ATPase activity assay, which showed that the highest activity was detected in the presence of mild detergents such as LDAO; moreover, high catalytic activity in the gel was already detected after the initial incubation period of 0.5 h. In contrast, HF<inf>1</inf>F<inf>0</inf> showed extremely low ATPase activity in harsher detergents such as TODC. The isolated fully functional enzyme will form the basis for future structural studies.

Original languageEnglish (US)
Pages (from-to)1-8
Number of pages8
JournalProtein Expression and Purification
StatePublished - Jun 8 2015



  • ATP hydrolysis
  • ATP synthase
  • Heliobacterium modesticaldum
  • In-gel function assay

ASJC Scopus subject areas

  • Biotechnology

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