Trypanosoma cruzi: Effect of Protein Kinase Inhibitors and Cytoskeletal Protein Organization and Expression on Host Cell Invasion by Amastigotes and Metacyclic Trypomastigotes

1998 ◽  
Vol 90 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Daniela O. Procópio ◽  
Solange da Silva ◽  
C.Casey Cunningham ◽  
Renato A. Mortara
Keyword(s):  
Protein Kinase ◽  
Trypanosoma Cruzi ◽  
Host Cell ◽  
Cell Invasion ◽  
Kinase Inhibitors ◽  
Protein Kinase Inhibitors ◽  
Cytoskeletal Protein ◽  
Host Cell Invasion ◽  
Metacyclic Trypomastigotes

scholarly journals Protein Kinase A Is Essential for Invasion of Plasmodium falciparum into Human Erythrocytes

mBio ◽  
2019 ◽  
Vol 10 (5) ◽  
Author(s):  
Mary-Louise Wilde ◽  
Tony Triglia ◽  
Danushka Marapana ◽  
Jennifer K. Thompson ◽  
Alexei A. Kouzmitchev ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Protein Kinase ◽  
Protein Kinase A ◽  
Host Cell ◽  
Cell Invasion ◽  
Blood Cells ◽  
Host Cell Invasion ◽  
Parasite Invasion ◽  
Content Type ◽  
Kinase A

ABSTRACT Understanding the mechanisms behind host cell invasion by Plasmodium falciparum remains a major hurdle to developing antimalarial therapeutics that target the asexual cycle and the symptomatic stage of malaria. Host cell entry is enabled by a multitude of precisely timed and tightly regulated receptor-ligand interactions. Cyclic nucleotide signaling has been implicated in regulating parasite invasion, and an important downstream effector of the cAMP-signaling pathway is protein kinase A (PKA), a cAMP-dependent protein kinase. There is increasing evidence that P. falciparum PKA (PfPKA) is responsible for phosphorylation of the cytoplasmic domain of P. falciparum apical membrane antigen 1 (PfAMA1) at Ser610, a cAMP-dependent event that is crucial for successful parasite invasion. In the present study, CRISPR-Cas9 and conditional gene deletion (dimerizable cre) technologies were implemented to generate a P. falciparum parasite line in which expression of the catalytic subunit of PfPKA (PfPKAc) is under conditional control, demonstrating highly efficient dimerizable Cre recombinase (DiCre)-mediated gene excision and complete knockdown of protein expression. Parasites lacking PfPKAc show severely reduced growth after one intraerythrocytic growth cycle and are deficient in host cell invasion, as highlighted by live-imaging experiments. Furthermore, PfPKAc-deficient parasites are unable to phosphorylate PfAMA1 at Ser610. This work not only identifies an essential role for PfPKAc in the P. falciparum asexual life cycle but also confirms that PfPKAc is the kinase responsible for phosphorylating PfAMA1 Ser610. IMPORTANCE Malaria continues to present a major global health burden, particularly in low-resource countries. Plasmodium falciparum, the parasite responsible for the most severe form of malaria, causes disease through rapid and repeated rounds of invasion and replication within red blood cells. Invasion into red blood cells is essential for P. falciparum survival, and the molecular events mediating this process have gained much attention as potential therapeutic targets. With no effective vaccine available, and with the emergence of resistance to antimalarials, there is an urgent need for the development of new therapeutics. Our research has used genetic techniques to provide evidence of an essential protein kinase involved in P. falciparum invasion. Our work adds to the current understanding of parasite signaling processes required for invasion, highlighting PKA as a potential drug target to inhibit invasion for the treatment of malaria.

scholarly journals Host cell invasion and oral infection by Trypanosoma cruzi strains of genetic groups TcI and TcIV from chagasic patients

2016 ◽  
Vol 9 (1) ◽  
Author(s):  
Fernando Yukio Maeda ◽  
Tatiana Mordente Clemente ◽  
Silene Macedo ◽  
Cristian Cortez ◽  
Nobuko Yoshida
Keyword(s):  
Trypanosoma Cruzi ◽  
Host Cell ◽  
Cell Invasion ◽  
Oral Infection ◽  
Host Cell Invasion ◽  
Genetic Groups

scholarly journals Expression and Cellular Localization of Molecules of the gp82 Family in Trypanosoma cruzi Metacyclic Trypomastigotes

2007 ◽  
Vol 75 (7) ◽  
pp. 3264-3270 ◽  
Author(s):  
Vanessa D. Atayde ◽  
Mauro Cortez ◽  
Renata Souza ◽  
José Franco da Silveira ◽  
Nobuko Yoshida
Keyword(s):  
Trypanosoma Cruzi ◽  
Host Cell ◽  
Cell Invasion ◽  
Cellular Localization ◽  
Polyclonal Antibodies ◽  
Microscopic Analysis ◽  
Cdna Clones ◽  
Host Cell Invasion ◽  
Two Dimensional Gel Electrophoresis ◽  
Surface Localization

ABSTRACT A member of the Trypanosoma cruzi gp82 family, expressed on metacyclic trypomastigote surface and identified by monoclonal antibody (MAb) 3F6, plays a key role in host cell invasion. Apart from the gp82 defined by MAb 3F6, no information is available on members of this protein family. From cDNA clones encoding gp82 proteins sharing 59.1% sequence identity, we produced the recombinant proteins J18 and C03, the former containing and the latter lacking the epitope for MAb 3F6. Polyclonal antibodies to J18 and C03 proteins were generated and used, along with MAb 3F6, to analyze the expression and cellular localization of gp82 family members in metacyclic forms of CL and G strains, which belong to highly divergent genetic groups. By two-dimensional gel electrophoresis and immunoblotting, molecules of 82 to 86 kDa, focusing at pH 4.6 to 5.4, and molecules of 72 to 88 kDa, focusing at pH 4.9 to 5.7, were visualized in CL and G strains, respectively. Flow cytometry and microscopic analysis revealed in both strains similar expression of MAb 3F6-reactive gp82 in live and permeabilized parasites, indicating its surface localization. The reaction of live parasites of both strains with anti-J18 antibodies was weaker than with MAb 3F6 and was increased by permeabilization. Anti-C03 antibodies bound predominantly to flagellar components in permeabilized G strain parasites, but in the CL strain the flagellum was not the preferential target for these antibodies. Host cell invasion of metacyclic forms was inhibited by J18 protein, as well as by MAb 3F6 and anti-J18 antibodies, but not by C03 protein or anti-C03 antibodies.

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