Abstract
The completion of the genome sequence of Trypanosoma cruzi has been followed by several studies of protein expression, with the long-term aim to obtain a complete picture of the parasite proteome. We report a proteomic analysis of an organellar cell fraction from T. cruzi CL Brener epimastigotes. A total of 396 proteins were identified by LC-MS/MS. Of these, 138 were annotated as hypothetical in the genome databases and the rest could be assigned to several metabolic and biosynthetic pathways, transport, and structural functions. Comparative analysis with a whole cell proteome study resulted in the validation of the expression of 173 additional proteins. Of these, 38 proteins previously reported in other stages were not found in the only large-scale study of the total epimastigote stage proteome. A selected set of identified proteins was analyzed further to investigate gene copy number, sequence variation, transmembrane domains, and targeting signals. The genes were cloned and the proteins expressed with a c-myc epitope tag in T. cruzi epimastigotes. Immunofluorescence microscopy revealed the localization of these proteins in different cellular compartments such as ER, acidocalcisome, mitochondrion, and putative cytoplasmic transport or delivery vesicles. The results demonstrate that the use of enriched subcellular fractions allows the detection of T. cruzi proteins that are undetected by whole cell proteomic methods.
| Original language | English |
|---|---|
| Pages (from-to) | 2735-2749 |
| Number of pages | 15 |
| Journal | Proteomics |
| Volume | 8 |
| Issue number | 13 |
| DOIs | |
| State | Published - Jul 1 2008 |
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Keywords
- Immunofluorescence
- Organelles
- Protein isoforms
- Trypanosoma cruzi
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Proteomics in Trypanosoma cruzi- Localization of novel proteins to various organelles. / Ferella, Marcela; Nilsson, Daniel; Darban, Hamid; Rodrigues, Claudia D; Bontempi, Esteban J.; Docampo, Roberto; Andersson, Björn.
In: Proteomics, Vol. 8, No. 13, 01.07.2008, p. 2735-2749.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Proteomics in Trypanosoma cruzi- Localization of novel proteins to various organelles
AU - Ferella, Marcela
AU - Nilsson, Daniel
AU - Darban, Hamid
AU - Rodrigues, Claudia D
AU - Bontempi, Esteban J.
AU - Docampo, Roberto
AU - Andersson, Björn
PY - 2008/7/1
Y1 - 2008/7/1
N2 - The completion of the genome sequence of Trypanosoma cruzi has been followed by several studies of protein expression, with the long-term aim to obtain a complete picture of the parasite proteome. We report a proteomic analysis of an organellar cell fraction from T. cruzi CL Brener epimastigotes. A total of 396 proteins were identified by LC-MS/MS. Of these, 138 were annotated as hypothetical in the genome databases and the rest could be assigned to several metabolic and biosynthetic pathways, transport, and structural functions. Comparative analysis with a whole cell proteome study resulted in the validation of the expression of 173 additional proteins. Of these, 38 proteins previously reported in other stages were not found in the only large-scale study of the total epimastigote stage proteome. A selected set of identified proteins was analyzed further to investigate gene copy number, sequence variation, transmembrane domains, and targeting signals. The genes were cloned and the proteins expressed with a c-myc epitope tag in T. cruzi epimastigotes. Immunofluorescence microscopy revealed the localization of these proteins in different cellular compartments such as ER, acidocalcisome, mitochondrion, and putative cytoplasmic transport or delivery vesicles. The results demonstrate that the use of enriched subcellular fractions allows the detection of T. cruzi proteins that are undetected by whole cell proteomic methods.
AB - The completion of the genome sequence of Trypanosoma cruzi has been followed by several studies of protein expression, with the long-term aim to obtain a complete picture of the parasite proteome. We report a proteomic analysis of an organellar cell fraction from T. cruzi CL Brener epimastigotes. A total of 396 proteins were identified by LC-MS/MS. Of these, 138 were annotated as hypothetical in the genome databases and the rest could be assigned to several metabolic and biosynthetic pathways, transport, and structural functions. Comparative analysis with a whole cell proteome study resulted in the validation of the expression of 173 additional proteins. Of these, 38 proteins previously reported in other stages were not found in the only large-scale study of the total epimastigote stage proteome. A selected set of identified proteins was analyzed further to investigate gene copy number, sequence variation, transmembrane domains, and targeting signals. The genes were cloned and the proteins expressed with a c-myc epitope tag in T. cruzi epimastigotes. Immunofluorescence microscopy revealed the localization of these proteins in different cellular compartments such as ER, acidocalcisome, mitochondrion, and putative cytoplasmic transport or delivery vesicles. The results demonstrate that the use of enriched subcellular fractions allows the detection of T. cruzi proteins that are undetected by whole cell proteomic methods.
KW - Immunofluorescence
KW - Organelles
KW - Protein isoforms
KW - Trypanosoma cruzi
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UR - http://www.scopus.com/inward/citedby.url?scp=47249134299&partnerID=8YFLogxK
U2 - 10.1002/pmic.200700940
DO - 10.1002/pmic.200700940
M3 - Article
VL - 8
SP - 2735
EP - 2749
JO - Proteomics
JF - Proteomics
SN - 1615-9853
IS - 13
ER -