Archives of Microbiology

, Volume 179, Issue 5, pp 368–376 | Cite as

Rhodobacter sphaeroides has a family II pyrophosphatase: comparison with other species of photosynthetic bacteria

  • Heliodoro Celis
  • Bernardo Franco
  • Silvia Escobedo
  • Irma Romero
Original Paper

Abstract

The cytoplasmic pyrophosphatase from Rhodobacter sphaeroides was purified and characterized. The enzyme is a homodimer of 64 kDa. The N-terminus was sequenced and used to obtain the complete pyrophosphatase sequence from the preliminary genome sequence of Rba. sphaeroides, showing extensive sequence similarity to family II or class C pyrophosphatases. The enzyme hydrolyzes only Mg-PPi and Mn-PPi with a K m of 0.35 mM for both substrates. It is not activated by free Mg 2+, in contrast to the cytoplasmic pyrophosphatase from Rhodospirillum rubrum, and it is not inhibited by NaF, methylendiphosphate, or imidodiphosphate. This work shows that Rba. sphaeroides and Rhodobacter capsulatus cytoplasmic pyrophosphatases belong to family II, in contrast to Rsp. rubrum, Rhodopseudomonas palustris, Rhodopseudomonas gelatinosa, and Rhodomicrobium vannielii cytoplasmic pyrophosphatases which should be classified as members of family I. This is the first report of family II cytoplasmic pyrophosphatases in photosynthetic bacteria and in a gram-negative organism.

Keywords

Photosynthetic bacteria Rhodobacter sphaeroides Rhodospirillum rubrum Rhodobacter capsulatus Family II inorganic pyrophosphatase 

Abbreviations

MDP

Methylendiphosphate

IDP

Imidodiphosphate

CAPS

3-[Cyclohexylamino]-1-propanesulfonic acid

Notes

Acknowledgments

This work was supported in part by grant IN 216401 from DGAPA. We thank the Molecular Biology Unit of the IFC for sequencing genes, and primer synthesis. The authors would like to thank Dr. Mark West for his assistance with protein purification and suggestions regarding the manuscript. We also thank Dr. Georges Dreyfus for critically reading the manuscript.

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Heliodoro Celis
    • 1
  • Bernardo Franco
    • 1
  • Silvia Escobedo
    • 1
  • Irma Romero
    • 2
  1. 1.Instituto de Fisiología CelularUniversidad Nacional Autónoma de MéxicoMéxico
  2. 2.Departamento de Bioquímica, Facultad de MedicinaUniversidad Nacional Autónoma de MéxicoMéxico

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