Proteomic Analysis of the Acidocalcisome, an Organelle Conserved From Bacteria to Human Cells

PLoS Pathog. 2014 Dec 11;10(12):e1004555. doi: 10.1371/journal.ppat.1004555. eCollection 2014 Dec.

Abstract

Acidocalcisomes are acidic organelles present in a diverse range of organisms from bacteria to human cells. In this study acidocalcisomes were purified from the model organism Trypanosoma brucei, and their protein composition was determined by mass spectrometry. The results, along with those that we previously reported, show that acidocalcisomes are rich in pumps and transporters, involved in phosphate and cation homeostasis, and calcium signaling. We validated the acidocalcisome localization of seven new, putative, acidocalcisome proteins (phosphate transporter, vacuolar H+-ATPase subunits a and d, vacuolar iron transporter, zinc transporter, polyamine transporter, and acid phosphatase), confirmed the presence of six previously characterized acidocalcisome proteins, and validated the localization of five novel proteins to different subcellular compartments by expressing them fused to epitope tags in their endogenous loci or by immunofluorescence microscopy with specific antibodies. Knockdown of several newly identified acidocalcisome proteins by RNA interference (RNAi) revealed that they are essential for the survival of the parasites. These results provide a comprehensive insight into the unique composition of acidocalcisomes of T. brucei, an important eukaryotic pathogen, and direct evidence that acidocalcisomes are especially adapted for the accumulation of polyphosphate.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Calcium Signaling / physiology
  • Cations / metabolism
  • Cells, Cultured
  • Homeostasis / physiology
  • Mass Spectrometry
  • Membrane Transport Proteins / metabolism*
  • Organelles / metabolism*
  • Phosphates / metabolism
  • Proteomics*
  • Protozoan Proteins / metabolism*
  • Trypanosoma brucei brucei / cytology*
  • Trypanosoma brucei brucei / metabolism

Substances

  • Cations
  • Membrane Transport Proteins
  • Phosphates
  • Protozoan Proteins