scholarly journals Molecular and Biochemical Characterization of a Type II Thioesterase From the Zoonotic Protozoan Parasite Cryptosporidium parvum

2019 ◽  
Vol 9 ◽  
Author(s):  
Fengguang Guo ◽  
Haili Zhang ◽  
Rana Eltahan ◽  
Guan Zhu
Keyword(s):  
Cryptosporidium Parvum ◽  
Biochemical Characterization ◽  
Protozoan Parasite ◽  
Type Ii

Biochemical characterization of a novel type-II VEGFR2 kinase inhibitor: Comparison of binding to non-phosphorylated and phosphorylated VEGFR2

2011 ◽  
Vol 19 (18) ◽  
pp. 5342-5351 ◽  
Author(s):  
Hidehisa Iwata ◽  
Shinichi Imamura ◽  
Akira Hori ◽  
Mark S. Hixon ◽  
Hiroyuki Kimura ◽  
...  
Keyword(s):  
Biochemical Characterization ◽  
Kinase Inhibitor ◽  
Type Ii

scholarly journals Characterization of INS-15, A Metalloprotease Potentially Involved in the Invasion of Cryptosporidium parvum

2019 ◽  
Vol 7 (10) ◽  
pp. 452 ◽  
Author(s):  
Xu ◽  
Guo ◽  
Li ◽  
Zhang ◽  
Wu ◽  
...  
Keyword(s):  
Cryptosporidium Parvum ◽  
Polyclonal Antibodies ◽  
Protozoan Parasite ◽  
Partial Reduction ◽  
E Coli ◽  
Severe Diarrhea ◽  
Domain I

Cryptosporidium parvum is a protozoan parasite that can cause moderate-to-severe diarrhea. Insulinase-like proteases (INS) are one of the largest protein families within the small proteome of the pathogen. However, their roles in C. parvum biology remain un-elucidated. In this study, a member of the protein family, INS-15 of C. parvum encoded by cgd3_4260, was cloned, expressed and characterized to understand its function. INS-15 and its domain I were expressed in Escherichia coli and polyclonal antibodies against the domain I and one specific polypeptide were prepared in rabbits. The role of INS-15 protein in the C. parvum invasion was preliminarily studied. Recombinant INS-15 protein and its domain I were successfully expressed in E. coli, together with various degraded products. The cgd3_4260 gene had a peak expression at 2 h of in vitro C. parvum culture, while the INS-15 protein was expressed in the mid-anterior region of sporozoites and the area of merozoites opposite to the nucleus. Anti-INS-15 domain I antibodies reduced the invasion of C. parvum sporozoites by over 40%. The anterior location of INS-15 in invasion stages and partial reduction of in vitro growth indicate that INS-15 plays some roles in the invasion or early development of C. parvum.

scholarly journals Structural and Biochemical Characterization of the Type II Fructose-1,6-bisphosphatase GlpX fromEscherichia coli

2008 ◽  
Vol 284 (6) ◽  
pp. 3784-3792 ◽  
Author(s):  
Greg Brown ◽  
Alexander Singer ◽  
Vladimir V. Lunin ◽  
Michael Proudfoot ◽  
Tatiana Skarina ◽  
...  
Keyword(s):  
Biochemical Characterization ◽  
Type Ii ◽  
Fromescherichia Coli ◽  
Fructose 1,6 Bisphosphatase

scholarly journals Characterization of an Intestinal Epithelial Cell Receptor Recognized by the Cryptosporidium parvum Sporozoite Ligand CSL

2001 ◽  
Vol 69 (3) ◽  
pp. 1661-1670 ◽  
Author(s):  
Rebecca C. Langer ◽  
Deborah A. Schaefer ◽  
Michael W. Riggs
Keyword(s):  
Cryptosporidium Parvum ◽  
Surface Protein ◽  
Protozoan Parasite ◽  
Mesenchymal Cell ◽  
Cell Receptor ◽  
Receptor Expression ◽  
Intestinal Epithelial ◽  
Protein Functions ◽  
Important Host

ABSTRACT The protozoan parasite Cryptosporidium parvum is a leading cause of diarrhea in humans and neonatal calves. The absence of approved parasite-specific drugs, vaccines, and immunotherapies for cryptosporidiosis relates in part to limited knowledge on the pathogenesis of zoite attachment and invasion. We recently reported that the C. parvum apical complex glycoprotein CSL contains a zoite ligand for intestinal epithelial cells which is defined by monoclonal antibody (MAb) 3E2. In the present study, the host cell receptor for CSL was characterized. For these studies, a panel of epithelial and mesenchymal cell lines was examined for permissiveness to C. parvum and the ability to bind CSL. Cells of epithelial origin were significantly more permissive and bound significantly greater quantities of CSL than cells of mesenchymal origin. Caco-2 intestinal cells were selected from the epithelial panel for further characterization of the CSL receptor. Immunoelectron microscopy demonstrated that CSL bound initially to the surface of Caco-2 cells and was rapidly internalized. The molecule bound by CSL was identified as an 85-kDa Caco-2 cell surface protein by radioimmunoprecipitation and CSL affinity chromatography. Sporozoite incubation with the isolated 85-kDa protein reduced binding of MAb 3E2. Further, attachment and invasion were significantly inhibited when sporozoites were incubated with the 85-kDa protein prior to inoculation onto Caco-2 cells. These observations indicate that the 85-kDa protein functions as a Caco-2 cell receptor for CSL. CSL also bound specifically to intestinal epithelium from calves, indicating receptor expression in a second important host species. Molecular characterization of the CSL receptor may lead to novel avenues for disrupting ligand-receptor interactions in the pathogenesis of C. parvum infection.

scholarly journals Toxoplasma gondii serine hydrolases regulate parasite lipid mobilization during growth and replication within the host

2020 ◽  
Author(s):  
Ouma Onguka ◽  
Brett M. Babin ◽  
Markus Lakemeyer ◽  
Ian T. Foe ◽  
Neri Amara ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Biochemical Characterization ◽  
Lipid Synthesis ◽  
Protozoan Parasite ◽  
List Type ◽  
Serine Hydrolases ◽  
Metabolizing Enzymes ◽  
Thiol Esters ◽  
Intracellular Protozoan Parasite

SummaryThe intracellular protozoan parasite Toxoplasma gondii must scavenge cholesterol and other lipids from the host to facilitate intracellular growth and replication. Enzymes responsible for neutral lipid synthesis have been identified but there is no evidence for enzymes that catalyze lipolysis of cholesterol esters and esterified lipids. Here we characterize several T. gondii serine hydrolases with esterase and thioesterase activities that were previously thought to be depalmitoylating enzymes. We find they do not cleave palmitoyl thiol esters but rather hydrolyze short chain lipid esters. Deletion of one of the hydrolases results in alterations in levels of multiple lipids species. We also identify small molecule inhibitors of these hydrolases and show that treatment of parasites results in phenotypic defects reminiscent of parasites exposed to excess cholesterol or oleic acid. Together, these data characterize enzymes necessary for processing lipids critical for infection and highlight the potential for targeting parasite hydrolases for therapeutic applications.HighlightsBioinformatic and biochemical characterization of T. gondii serine hydrolases reveals substrate preference between enzymes with similar catalytic foldT. gondii serine hydrolases previously thought to be depalmitoylases are lipid metabolizing enzymesT. gondii lipid metabolism pathways utilize enzymes that are viable therapeutic targets

scholarly journals Antibodies Raised against Bcvir15, an Extrachromosomal Double-Stranded RNA-Encoded Protein from Babesia canis, Inhibit the In Vitro Growth of the Parasite

2003 ◽  
Vol 71 (3) ◽  
pp. 1056-1067 ◽  
Author(s):  
P. Drakulovski ◽  
B. Carcy ◽  
K. Moubri ◽  
C. Carret ◽  
D. Depoix ◽  
...  
Keyword(s):  
Biochemical Characterization ◽  
Protozoan Parasite ◽  
Inhibitory Effect ◽  
Open Reading Frames ◽  
Babesia Canis ◽  
In Vitro Growth ◽  
Viral Origin ◽  
Double Stranded Rna

ABSTRACT As part of a search for homologous members of the Plasmodium falciparum Pf60 multigene family in the intraerythrocytic protozoan parasite Babesia canis, we report here the characterization of a cDNA of 1,115 bp, which was designated Bcvir for its potential viral origin. The Bcvir cDNA contained two overlapping open reading frames (ORFs) (ORF1 from nucleotide [nt] 61 to 486 and ORF2 from nt 417 to 919), where Bcvir15, the deduced ORF1 peptide (M1 to I141), is the main expressed product. The Bcvir cDNA was derived from an extrachromosomal dsRNA element of 1.2 kbp that was always found associated with a double-stranded RNA (dsRNA) of 2.8 kbp by hybridization, and no copy of this cDNA sequence was found in B. canis genomic DNA. Biochemical characterization of Bcvir15, by using polyclonal rabbit sera directed against recombinant proteins, indicated that it is a soluble protein which remained associated with the cytoplasm of the B. canis merozoite. Interestingly, purified immunoglobulins from the anti-glutathione S-transferase-Bcvir15 (at a concentration of 160 μg/ml) induced 50% inhibition of the in vitro growth of B. canis, and the inhibitory effect was associated with morphological damage of the parasite. Our data suggest that the extrachromosomal dsRNA-encoded Bcvir15 protein might interfere with the intracellular growth of the parasite rather than with the process of invasion of the host cell by the merozoite. Epitope mapping of Bcvir15 identified three epitopes that might be essential for the function of the protein.

scholarly journals Biochemical characterization of enoyl reductase involved in Type II fatty acid synthesis in the intestinal coccidiumEimeria tenella(PhylumApicomplexa)

2007 ◽  
Vol 272 (2) ◽  
pp. 238-244 ◽  
Author(s):  
Xiaomin Cai ◽  
A. Lorraine Fuller ◽  
Larry R. McDougald ◽  
Xiangshi Tan ◽  
Jianping Cai ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Synthesis ◽  
Biochemical Characterization ◽  
Acid Synthesis ◽  
Type Ii ◽  
Enoyl Reductase

Purification and partial biochemical characterization of a membrane-bound type II-like α-glucosidase from the yeast morphotype of Sporothrix schenckii

2011 ◽  
Vol 101 (2) ◽  
pp. 313-322 ◽  
Author(s):  
Blanca I. Torres-Rodríguez ◽  
Karina Flores-Berrout ◽  
Julio C. Villagómez-Castro ◽  
Everardo López-Romero
Keyword(s):  
Biochemical Characterization ◽  
Sporothrix Schenckii ◽  
Type Ii ◽  
Membrane Bound
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