16 research outputs found
Identification of Contractile Vacuole Proteins in Trypanosoma cruzi
Contractile vacuole complexes are critical components of cell volume regulation
and have been shown to have other functional roles in several free-living
protists. However, very little is known about the functions of the contractile
vacuole complex of the parasite Trypanosoma cruzi, the
etiologic agent of Chagas disease, other than a role in osmoregulation.
Identification of the protein composition of these organelles is important for
understanding their physiological roles. We applied a combined proteomic and
bioinfomatic approach to identify proteins localized to the contractile vacuole.
Proteomic analysis of a T. cruzi fraction enriched for
contractile vacuoles and analyzed by one-dimensional gel electrophoresis and
LC-MS/MS resulted in the addition of 109 newly detected proteins to the group of
expressed proteins of epimastigotes. We also identified different peptides that
map to at least 39 members of the dispersed gene family 1 (DGF-1) providing
evidence that many members of this family are simultaneously expressed in
epimastigotes. Of the proteins present in the fraction we selected several
homologues with known localizations in contractile vacuoles of other organisms
and others that we expected to be present in these vacuoles on the basis of
their potential roles. We determined the localization of each by expression as
GFP-fusion proteins or with specific antibodies. Six of these putative proteins
(Rab11, Rab32, AP180, ATPase subunit B, VAMP1, and phosphate transporter)
predominantly localized to the vacuole bladder. TcSNARE2.1, TcSNARE2.2, and
calmodulin localized to the spongiome. Calmodulin was also cytosolic. Our
results demonstrate the utility of combining subcellular fractionation,
proteomic analysis, and bioinformatic approaches for localization of organellar
proteins that are difficult to detect with whole cell methodologies. The CV
localization of the proteins investigated revealed potential novel roles of
these organelles in phosphate metabolism and provided information on the
potential participation of adaptor protein complexes in their biogenesis
Pulse Gradient Spin Echo Measurement of Flow Dynamics in a Porous Structure: NMR Spectral Analysis of Motional Correlations
Global gene response in saccharomyces cerevisiae exposed to silver nanoparticles
Silver nanoparticles (AgNPs), exhibiting a broad size range and morphologies with highly reactive facets, which are widely applicable in real-life but not fully verified for biosafety and ecotoxicity, were subjected to report transcriptome profile in yeast Saccharomyces cerevisiae. A large number of genes accounted for similar to 3% and similar to 5% of the genome affected by AgNPs and Ag-ions, respectively. Principal component and cluster analysis suggest that the different physical forms of Ag were the major cause in differential expression profile. Among 90 genes affected by both AgNPs and Ag-ions, metalloprotein mediating high resistance to copper (CUP1-1 and CUP1-2) were strongly induced by AgNPs (similar to 45-folds) and Ag-ions (similar to 22-folds), respectively. A total of 17 genes, responsive to chemical stimuli, stress, and transport processes, were differentially induced by AgNPs. The differential expression was also seen with Ag-ions that affected 73 up- and 161 down-regulating genes, and most of these were involved in ion transport and homeostasis. This study provides new information on the knowledge for impact of nanoparticles on living microorganisms that can be extended to other nanoparticles