Proteomic characterization of the subpellicular cytoskeleton of Toxoplasma gondii tachyzoites

León, Carmen T. Gómez de; Martín, Rubén Darío Díaz; Hernández, Guillermo Mendoza; Pozos, Sirenia González; Ambrosio, Javier R.; Flores, Ricardo Mondragón

doi:10.1016/j.jprot.2014.03.008

Cited by 23 publications

(18 citation statements)

References 69 publications

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“…Interestingly, a network of intermediate filament‐like proteins, the alveolins, underlies the cytoplasmic face of the IMC and might interact with PAP1 . Supporting this hypothesis, PAP1 has been found in the proteome of the subpellicular cytoskeleton of T. gondii and the annuli are resistant to oryzalin‐mediated microtubule depolymerization. Regarding PAP2, its sequence is predicted to include ARM repeats and a coiled‐coil domain.…”

Section: Discussionmentioning

confidence: 99%

The roles of Centrin 2 and Dynein Light Chain 8a in apical secretory organelles discharge of Toxoplasma gondii

Lentini

Dubois

Maco

et al. 2019

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To efficiently enter host cells, apicomplexan parasites such as Toxoplasma gondii rely on an apical complex composed of tubulin‐based structures as well as two sets of secretory organelles named micronemes and rhoptries. The trafficking and docking of these organelles to the apical pole of the parasite is crucial for the discharge of their contents. Here, we describe two proteins typically associated with microtubules, Centrin 2 (CEN2) and Dynein Light Chain 8a (DLC8a), that are required for efficient host cell invasion. CEN2 localizes to four different compartments, and remarkably, conditional depletion of the protein occurs in stepwise manner, sequentially depleting the protein pools from each location. This phenomenon allowed us to discern the essential function of the apical pool of CEN2 for microneme secretion, motility, invasion and egress. DLC8a localizes to the conoid, and its depletion also perturbs microneme exocytosis in addition to the apical docking of the rhoptry organelles, causing a severe defect in host cell invasion. Phenotypic characterization of CEN2 and DLC8a indicates that while both proteins participate in microneme secretion, they likely act at different steps along the cascade of events leading to organelle exocytosis.

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Section: Discussionmentioning

confidence: 99%

The roles of Centrin 2 and Dynein Light Chain 8a in apical secretory organelles discharge of Toxoplasma gondii

Lentini

Dubois

Maco

et al. 2019

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“…Cover slides were mounted with vectashield mounting medium (Vector Laboratories, Inc., Burlingame, CA, USA) and observed in a fluorescence microscope (Axioskop 2 mot plus, Carl Zeiss, Germany). The images were recorded in an Axiocam HR digital camera and processed with the Axiovision software (Carl Zeiss, Version 4.8), (Gomez de Leon et al 2014). …”

Section: Fluorescence Microscopymentioning

confidence: 99%

“…Detection of PP2C in L. major promastigotes was achieved by electron microscopy (Gomez de Leon et al 2014). Briefly, promastigotes were washed with PBS and then fixed with a mixture of 4% paraformaldehyde and 0·1% glutaraldehyde in PBS for 1 h at RT.…”

Section: Immunolocalization Of Pp2c By Immune Electron Microscopy (Iem)mentioning

confidence: 99%

Protein phosphatase PP2C in the flagellum of Leishmania major: cloning and characterization

et al. 2017

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The main goal of this work consisted in cloning, purifying and characterizing a protein phosphatase 2C (PP2C) from promastigotes of Leishmania major. The gene was cloned and amplified by PCR using specific oligonucleotides and the recombinant protein was purified by affinity chromatography. The peak with maximal protein concentration was analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and revealed a protein of 44·9 kDa with PP2C activity. This activity was dependent on divalent cations (Mg +2 and Mn +2 ) and was optimal at pH of 8·5, using phosphothreonine as the substrate. Sanguinarine inhibited the activity of the recombinant LmPP2C, while protein tyrosine phosphatase inhibitors had no effect. The recombinant LmPP2C was used to generate polyclonal antibodies. These antibodies recognized a protein of 44·9 kDa in different Leishmania species; the LmPP2C was localized in the flagellar pocket and the flagellum of promastigotes.

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“…In addition to these scaffolding proteins and the anchoring components of the motility apparatus that reside in the IMC, many additional proteins are known to occupy the pellicle or IMC domain butmost have yet to be characterized. [20][21][22].The importance of pellicle architecture in structurally supporting normal parasite motility has been demonstrated by several studies in which disruption of pellicle components has led to altered gliding behavior [23][24][25][26].…”

Section: Apicomplexan Architecturementioning

confidence: 99%

Gliding motility in apicomplexan parasites

Heintzelman

2015

Seminars in Cell & Developmental Biology

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Proteomic characterization of the subpellicular cytoskeleton of Toxoplasma gondii tachyzoites

Cited by 23 publications

References 69 publications

The roles of Centrin 2 and Dynein Light Chain 8a in apical secretory organelles discharge of Toxoplasma gondii

The roles of Centrin 2 and Dynein Light Chain 8a in apical secretory organelles discharge of Toxoplasma gondii

Protein phosphatase PP2C in the flagellum of Leishmania major: cloning and characterization

Gliding motility in apicomplexan parasites

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