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A giant phosphoprotein localized at the spongiome region of Crithidia luciliae thermophila
A giant phosphoprotein localized at the spongiome region of Crithidia luciliae thermophila
- Source :
- The Journal of eukaryotic microbiology. 47(6)
- Publication Year :
- 2000
-
Abstract
- A giant protein with an apparent molecular mass of 2,300-kDa was identified in the Triton X-100 soluble fraction of Crithidia luciliae thermophila. Polyclonal antibody raised against this protein reacted by immunoblot analysis with proteins of similar molecular mass in Crithidia fasciculata and Crithidia oncopelti. In addition, the antibody immunoprecipitates the protein either after in vivo phosphorylation with [32P]orthophosphoric acid or after metabolically labeling with [35S]methionine. Indirect immunofluorescence microscopy analysis performed either with fixed or with live parasites showed a single fluorescent spot at the level of the flagellar pocket region. Immunogold electron microscopy of thin sections of the parasite revealed that the antigen is localized at a restricted area of the spongiome, between the contractile vacuole and the flagellar pocket. Furthermore, Triton X-114 phase separation of whole cell membrane proteins, metabolically labeled with [35S]methionine, demonstrated that the giant protein remains in the aqueous phase. These results indicate that this phosphoprotein behaves as a peripheral membrane protein localized at the spongiome region, suggesting that it might be involved in the osmoregulatory process.
- Subjects :
- Glycosylation
Protozoan Proteins
Antigens, Protozoan
Microbiology
Epitopes
Crithidia
Crithidia luciliae
Animals
Phosphorylation
Fluorescent Antibody Technique, Indirect
Microscopy, Immunoelectron
Crithidia fasciculata
Molecular mass
biology
Peripheral membrane protein
Immunogold labelling
biology.organism_classification
Phosphoproteins
Contractile vacuole
Molecular Weight
Biochemistry
Membrane protein
Microscopy, Fluorescence
Solubility
Phosphoprotein
Vacuoles
Protein Processing, Post-Translational
Subjects
Details
- ISSN :
- 10665234
- Volume :
- 47
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- The Journal of eukaryotic microbiology
- Accession number :
- edsair.doi.dedup.....240adbdc9e81bc7ca543fe46e2024637