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1 November 2000 A Giant Phosphoprotein Localized at the Spongiome Region of Crithidia luciliae thermophila
MUNIRA M. A. BAQUI, NADIA DE MORAES, REGINA V. MILDER, JULIO PUDLES
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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.

MUNIRA M. A. BAQUI, NADIA DE MORAES, REGINA V. MILDER, and JULIO PUDLES "A Giant Phosphoprotein Localized at the Spongiome Region of Crithidia luciliae thermophila," The Journal of Eukaryotic Microbiology 47(6), 532-537, (1 November 2000). https://doi.org/10.1111/j.1550-7408.2000.tb00086.x
Received: 21 February 2000; Accepted: 6 June 2000; Published: 1 November 2000
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KEYWORDS
Contractile vacuole
giant protein
peripheral membrane proteins
phosphorylation
trypanosomatid
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