Characterization of Farnesylated Protein Tyrosine Phosphatase TcPRL-1 from
            <i>Trypanosoma cruzi</i>

Cuevas, Ileana; Rohloff, Peter; Sánchez, Daniel O.; Docampo, Roberto

doi:10.1128/ec.4.9.1550-1561.2005

Cited by 32 publications

(26 citation statements)

References 47 publications

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“…In vitro prenylation reactions were performed as described previously (Cuevas et al, 2005) Ca 2+ uptake by digitonin-permeabilized epimastigotes was measured using the fluorescence indicator Calcium Green-5N, as described previously (Huang et al, 2013). Acidification of digitonin-permeabilized cells was followed by measuring the changes in the fluorescence of Acridine Orange in a fluorometer, as described previously (Docampo et al, 1995).…”

Section: In Vitro Prenylationmentioning

confidence: 99%

Rab32 is essential for maintaining functional acidocalcisomes, and for growth and infectivity of Trypanosoma cruzi

Niyogi

Jiménez

Girard-Dias

et al. 2015

Journal of Cell Science

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The contractile vacuole complex (CVC) of Trypanosoma cruzi, the etiologic agent of Chagas disease, collects and expels excess water as a mechanism of regulatory volume decrease after hyposmotic stress; it also has a role in cell shrinking after hyperosmotic stress. Here, we report that, in addition to its role in osmoregulation, the CVC of T. cruzi has a role in the biogenesis of acidocalcisomes. Expression of dominant-negative mutants of the CVC-located small GTPase Rab32 (TcCLB.506289.80) results in lower numbers of lesselectron-dense acidocalcisomes, lower content of polyphosphate, lower capacity for acidocalcisome acidification and Ca 2+ uptake that is driven by the vacuolar proton pyrophosphatase and the Ca 2+ -ATPase, respectively, as well as less-infective parasites, revealing the role of this organelle in parasite infectivity. By using fluorescence, electron microscopy and electron tomography analyses, we provide further evidence of the active contact of acidocalcisomes with the CVC, indicating an active exchange of proteins between the two organelles.

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Section: In Vitro Prenylationmentioning

confidence: 99%

Rab32 is essential for maintaining functional acidocalcisomes, and for growth and infectivity of Trypanosoma cruzi

Niyogi

Jiménez

Girard-Dias

et al. 2015

Journal of Cell Science

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“…All of these four T. cruzi PRL homologs contain the C-terminal CaaX sequence ending with Met, and thus are predicted to be prenylated similarly to TcPRL-1, which has been reported to be farnesylated [27]. Two T. cruzi DNAJ proteins (Tcj2 and Tcj4) that have previously been reported also contain the CaaX sequences [36].…”

Section: Substrates For the Putative T Cruzi Pggt-i In T Cruzi Cellsmentioning

confidence: 99%

“…Two proteins in T. cruzi, TcRho1 GTPase [26] and TcPRL-1 protein tyrosine phosphatase [27] have so far been shown to be farnesylated. Here we describe the cloning of a putative PGGT-I β subunit and co-expression of this gene in combination with the T. cruzi PFT α gene to yield a functional PGGT-I.…”

Section: Introductionmentioning

confidence: 99%

Protein geranylgeranyltransferase-I of Trypanosoma cruzi

Yokoyama

Gillespie

Voorhis

et al. 2008

Molecular and Biochemical Parasitology

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Protein geranylgeranyltransferase type I (PGGT-I) and protein farnesyltransferase (PFT) occur in many eukaryotic cells. Both consist of two subunits, the common αsubunit and a distinct β subunit. In the gene database of protozoa Trypanosoma cruzi, the causative agent of Chagas' disease, a putative protein that consists of 401 amino acids with ∼20% amino acid sequence identity to the PGGT-I β of other species was identified, cloned, and characterized. Multiple sequence alignments show that the T. cruzi ortholog contains all three of the zinc-binding residues and several residues uniquely conserved in the β subunit of PGGT-I. Co-expression of this protein and the α subunit of T. cruzi PFT in Sf9 insect cells yielded a dimeric protein that forms a tight complex selectively with [ 3 H] geranylgeranyl pyrophosphate, indicating a key characteristic of a functional PGGT-I. Recombinant T. cruzi PGGT-I ortholog showed geranylgeranyltransferase activity with distinct specificity toward the C-terminal CaaX motif of protein substrates compared to that of the mammalian PGGT-I and T. cruzi PFT. Most of the CaaX-containing proteins with X=Leu are good substrates of T. cruzi PGGT-I, and those with X=Met are substrates for both T. cruzi PFT and PGGT-I, whereas unlike mammalian PGGT-I, those with X=Phe are poor substrates for T. cruzi PGGT-I. Several candidates for T. cruzi PGGT-I or PFT substrates containing the C-terminal CaaX motif are found in the T. cruzi gene database. Among five C-terminal peptides of those tested, a peptide of a Ras-like protein ending with CVLL was selectively geranylgeranylated by T. cruzi PGGT-I. Other peptides with CTQQ (Tcj2 DNAJ protein), CAVM (TcPRL-1 protein tyrosine phosphatase), CHFM (a small GTPase like protein), and CQLF (TcRho1 GTPase) were specific substrates for T. cruzi PFT but not for PGGT-I. The mRNA and protein of the T. cruzi PGGT-I β ortholog were detected in three life-cycle stages of T. cruzi. Cytosol fractions from trypomastigotes (infectious mammalian stage) and epimastigotes (insect stage) were shown to contain levels of PGGT-I activity that are ∼100-fold lower than PFT activity. The CaaX mimetics known as PGGT-I inhibitors show very low potency against T. cruzi PGGT-I compared to the mammalian enzyme, suggesting the potential to develop selective inhibitors against the parasite enzyme.

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“…These include, for example, a small GTPase and a protein tyrosine phosphatase from T. cruzi (57,58) and two small GTPases from T. brucei (59,60). A bioinformatic approach, a search of potential prenylated proteins (those containing a cysteine at the À4 position from the C terminus) from T. brucei, yields 190 open reading frames of .50 amino acids.…”

Section: Pftismentioning

confidence: 99%

Thematic review series: Lipid Posttranslational Modifications. Fighting parasitic disease by blocking protein farnesylation

Eastman

Buckner

Yokoyama

et al. 2006

Journal of Lipid Research

103

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Protein farnesylation is a form of posttranslational modification that occurs in most, if not all, eukaryotic cells. Inhibitors of protein farnesyltransferase (PFTIs) have been developed as anticancer chemotherapeutic agents. Using the knowledge gained from the development of PFTIs for the treatment of cancer, researchers are currently investigating the use of PFTIs for the treatment of eukaryotic pathogens. This "piggy-back" approach not only accelerates the development of a chemotherapeutic agent for protozoan pathogens but is also a means of mitigating the costs associated with de novo drug design. PFTIs have already been shown to be efficacious in the treatment of eukaryotic pathogens in animal models, including both Trypanosoma brucei, the causative agent of African sleeping sickness, and Plasmodium falciparum, one of the causative agents of malaria. Here, current evidence and progress are summarized that support the targeting of protein farnesyltransferase for the treatment of parasitic diseases. Parasitic diseases continue to have a major impact on morbidity and mortality in tropical and subtropical regions. Among these, malaria causes $300 million infections annually, with 1-3 million deaths occurring in Africa (1). The emergence and spread of parasites resistant to existing antimalarial agents is largely responsible for the recent increase in malaria-related mortality. Another reemerging disease is African sleeping sickness (African trypanosomiasis), with an estimated 50,000 deaths in 2002 (1). The increasing burden of these diseases, along with the inadequacies of current drugs for African sleeping sickness in terms of safety, efficacy, and ease of administration, have led investigators to seek new chemotherapeutic agents (2, 3). Among the current drug targets under study are enzymes involved in protein prenylation, or the posttranslational modification of proteins by the covalent modification by isoprenyl lipids, C15 farnesyl and C20 geranylgeranyl (4-7). The isoprenyl lipid modification of proteins has been shown to be critical for various cellular activities in mammals and yeast, including proliferation and apoptosis (8, 9). Growth of the protozoan parasites has been shown to be severely impaired by the inhibition of protein farnesylation compared with mammalian cells, suggesting high potential of the enzyme protein farnesyltransferase (PFT) as an antiparasitic drug target (5, 10-13).The isoprenoid synthesis pathway from mevalonic acid in many eukaryotes, including trypanosomatids (or deoxyxylulose in Apicomplexa, including Plasmodium and Toxoplasma, and plants) is essential for the production of sterols, dolichol, ubiquinone, and other isoprene derivatives in many eukaryotic cells. Indeed, these pathways have been the study of recent efforts to develop other antiparasitic chemotherapeutic agents, especially the targeting of isoprenoid pyrophosphate synthesis by nitrogen-containing bisphosphonates (14-16). Organisms belonging to the group Apicomplexa contain the nonmevalonate pathway of iso...

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Characterization of Farnesylated Protein Tyrosine Phosphatase TcPRL-1 from Trypanosoma cruzi

Cited by 32 publications

References 47 publications

Rab32 is essential for maintaining functional acidocalcisomes, and for growth and infectivity of Trypanosoma cruzi

Rab32 is essential for maintaining functional acidocalcisomes, and for growth and infectivity of Trypanosoma cruzi

Protein geranylgeranyltransferase-I of Trypanosoma cruzi

Thematic review series: Lipid Posttranslational Modifications. Fighting parasitic disease by blocking protein farnesylation

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