Identification of organelles in bacteria similar to acidocalcisomes of unicellular eukaryotes

Manfredo Seufferheld, Mauricio C F Vieira, Felix A. Ruiz, Claudia D Rodrigues, Silvia N J Moreno, Roberto Docampo

Research output: Contribution to journalArticle

112 Citations (Scopus)

Abstract

Acidocalcisomes are acidic calcium storage compartments described in several unicellular eukaryotes, including trypanosomatid and apicomplexan parasites, algae, and slime molds. In this work, we report that the volutin granules of Agrobacterium tumefaciens possess properties similar to the acidocalcisomes. Transmission electron microscopy revealed that each intracellular granule was surrounded by a membrane. X-ray microanalysis of the volutin granules showed large amounts of phosphorus, magnesium, potassium, and calcium. Calcium in the volutin granules increased when the bacteria were incubated at high extracellular calcium concentration. Immunofluorescence and immunoelectron microscopy, using antisera raised against peptide sequences conserved in the A. tumefaciens proton pyrophosphatase, indicated localization in intracellular vacuoles. Purification of the volutin granules using iodixanol density gradients indicated a preferential localization of the pyrophosphatase activity in addition to high concentrations of phosphate, pyrophosphate, short- and long-chain polyphosphate, but lack of markers of the plasma membrane. The pyrophosphatase activity was potassium-insensitive and inhibited by the pyrophosphate analogs, amynomethylenediphosphonate and imidodiphosphate, by dicyclohexylcarbodiimide, and by the thiol reagent N-ethylmaleimide. Polyphosphate was also localized to the volutin granules by 4′, 6′-diamino-2-phenylindole staining. The organelles were acidic, as demonstrated by staining with LysoSensor blue DND-167, a dye especially used to detect very acidic compartments in cells, and cycloprodigiosin, a compound isolated from a marine bacterium that has been shown to uncouple proton pyrophosphatase activity acting as a chloride/proton symport. The results suggest that acidocalcisomes arose before the prokaryotic and eukaryotic lineages diverged.

Original languageEnglish
Pages (from-to)29971-29978
Number of pages8
JournalJournal of Biological Chemistry
Volume278
Issue number32
DOIs
StatePublished - Aug 8 2003
Externally publishedYes

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Pyrophosphatases
Eukaryota
Organelles
Bacteria
Calcium
Protons
Polyphosphates
Agrobacterium tumefaciens
Potassium
Staining and Labeling
Dicyclohexylcarbodiimide
Electron Probe Microanalysis
Sulfhydryl Reagents
Ethylmaleimide
Immunoelectron Microscopy
Conserved Sequence
Ion Transport
Cell membranes
Algae
Vacuoles

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of organelles in bacteria similar to acidocalcisomes of unicellular eukaryotes. / Seufferheld, Manfredo; Vieira, Mauricio C F; Ruiz, Felix A.; Rodrigues, Claudia D; Moreno, Silvia N J; Docampo, Roberto.

In: Journal of Biological Chemistry, Vol. 278, No. 32, 08.08.2003, p. 29971-29978.

Research output: Contribution to journalArticle

Seufferheld, Manfredo ; Vieira, Mauricio C F ; Ruiz, Felix A. ; Rodrigues, Claudia D ; Moreno, Silvia N J ; Docampo, Roberto. / Identification of organelles in bacteria similar to acidocalcisomes of unicellular eukaryotes. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 32. pp. 29971-29978.
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