Hydrodynamic characterization of the SufBC and SufCD complexes and their interaction with fluorescent adenosine nucleotides

Arsen Petrovic, Colin T. Davis, Kaveri Rangachari, Barbara Clough, R. J.M. Wilson, John F. Eccleston

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28 Citations (Scopus)

Abstract

Bacteria, as well as the plastid organelles of algae and higher plants, utilize proteins of the suf operon. These are involved in Fe-S cluster assembly, particularly under conditions of iron limitation or oxidative stress. Genetic experiments in some organisms found that the ATPase SufC is essential, though its role in Fe-S biogenesis remains unclear. To ascertain how interactions with other individual Suf proteins affect the activity of SufC we coexpressed it with either SufB or SufD from Thermotoga maritima and purified the resulting SufBC and SufCD complexes. Analytical ultracentrifuge and multiangle light-scattering measurements showed that the SufBC complex exists in solution as the tetrameric SufB2C2 species, whereas SufCD exists as an equilibrium mixture of SufCD and SufC2D2. Transient kinetic studies of the complexes were made using fluorescent 2′(3′)-O-(N- methylanthraniloyl-(mant) analogues of ATP and ADP. Both SufBC and SufCD bound mantATP and mantADP much more tightly than does SufC alone. Compared to the cleavage step of the mantATPase of SufC alone, that of SufBC was accelerated 180-fold and that of SufCD only fivefold. Given that SufB and SufD have 20% sequence identity and similar predicted secondary structures, the different hydrodynamic properties and kinetic mechanisms of the two complexes are discussed. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish
Pages (from-to)1264-1274
Number of pages11
JournalProtein Science
Volume17
Issue number7
DOIs
Publication statusPublished - Jul 2008

Keywords

  • ATPase
  • Hydrodynamics
  • Kinetic mechanism
  • Suf proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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