Abstract：
:Most of the signaling pathways are regulated by reversible phosphorylation-dephosphorylation which involves enzymes- kinases and phosphatases. Current knowledge about the protein phosphatases in parasites like Trypanosoma and Leishmania is very minimal despite their enormousity. In present study, full length ORF of Leishmania donovani PP2C was cloned into expression vector followed by purification and molecular weight determination using Ni-NTA affinity and gel giltration chromatography respectively. Purified LdPP2C was found to be enzymatically active, while inhibition study suggested that sanguinarine acts as a non-competitive inhibitor. CD and fluorescence spectroscopy results indicated towards an adequate protein conformation from pH 3.5 to 8.5. The quenching constant (Ksv) and free energy (ΔG) of LdPP2C was found to be 11.1 ± 0.2 mM-1 and 2.0 ± 1.1 kcal mol-1 in presence of acrylamide and urea respectively. The protein was found to elicit the innate immune functions through upregulation of pro-inflammatory cytokines (TNF-α and IL-6) as well as nitric oxide generation. Simultaneously, these cytokines were found to be fairly higher in protein treated cells as compared to untreated cells at transcript level too. These observations advocate that LdPP2C generates a pro-inflammatory environment in macrophages and hence plays important role in immunomodulation. Computational modelling showed similar three-dimensional structure and metal binding sites present in other member of PP2C subfamily, while docking studies revealed its interaction with substrate as well as its specific inhibitor. Our study has provided first time reports on enzyme kinetics, structural features and immune response inside the host macrophage of metal-dependent protein phosphatases from a trypanosomatid parasite.
journal_name
Mol Biochem Parasitoljournal_title
Molecular and biochemical parasitologyauthors
Jakkula P,Qureshi R,Iqbal A,Sagurthi SR,Qureshi IAdoi
10.1016/j.molbiopara.2018.06.005subject
Has Abstractpub_date
2018-07-01 00:00:00pages
37-49eissn
0166-6851issn
1872-9428pii
S0166-6851(18)30047-1journal_volume
223pub_type
杂志文章abstract：:The variant surface glycoproteins (VSGs) of both metacyclic and bloodstream forms of Trypanosoma congolense are shown to be anchored to the plasma membrane through a glycolipid similar to that found in Trypanosoma brucei. Release of soluble VSG from both metacyclic and bloodstream forms is associated with the exposure...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(87)90045-4
更新日期：1987-01-15 00:00:00
abstract：:Six discrete phylogenetic lineages were recently identified in Trypanosoma cruzi, on the basis of multilocus enzyme electrophoresis and random amplified polymorphic DNA (RAPD) characterisation. The objective of the present study was to develop specific PCR-based markers for the identification of each of the six lineag...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(00)00302-9
更新日期：2000-11-01 00:00:00
abstract：:The livers of chickens maintained on a diet containing an effective in vivo level (70 ppm) of arprinocid, an anticoccidial agent, were found to contain 0.64 ppm of this drug and 0.33 ppm of a metabolite, arprinocid-1-N-oxide. When arprinocid was tested against Eimeria tenella in cultures of chick kidney epithelial cel...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(80)90051-1
更新日期：1980-10-01 00:00:00
abstract：:Post-transcriptional regulation is a key feature controlling gene expression in the protozoan parasite Leishmania. The nine-nucleotide paraflagellar rod regulatory element (PRE) in the 3'UTR of Leishmania mexicana PFR2 is both necessary and sufficient for the observed 10-fold higher level of PFR2 mRNA in promastigotes...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2007.10.001
更新日期：2008-01-01 00:00:00
abstract：:Erythrocyte invasion by malaria parasites requires specific receptor-ligand interactions. Plasmodium vivax and Plasmodium knowlesi are completely dependent on binding the Duffy blood group antigen to invade human erythrocytes. P. knowlesi invades rhesus erythrocytes by multiple pathways using the Duffy antigen as well...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(02)00017-8
更新日期：2002-04-30 00:00:00
abstract：:Growth inhibition and radioisotope incorporation studies with allopurinol (4-hydroxypyrazolo(3,4-d)pyrimidine) have shown that the African trypanosomes are biologically and biochemically similar to Leishmania spp. and Trypanosoma cruzi with respect to their response to this compound. These organisms, as a group, share...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(80)90001-8
更新日期：1980-04-01 00:00:00
abstract：:The number and expression of some of the large ribosomal RNA (rRNA) gene classes present in the human malaria parasite Plasmodium falciparum was determined. Southern blot analyses, using the 5.8S rRNA coding region as a marker, indicate that the P. falciparum genome contains at least 5 distinct subclasses of large rRN...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(90)90211-4
更新日期：1990-01-01 00:00:00
abstract：:Localization of thymidylate synthase protein in Trichinella spiralis and Caenorhabditis elegans development was followed with the use of confocal microscopy, revealing similar expression patterns in both nematode species. In T. spiralis premature muscle larvae and C. elegans dauer, L3 and L4 larvae, thymidylate syntha...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2012.02.002
更新日期：2012-05-01 00:00:00
abstract：:The major surface antigen (30 kDa) of Brugia pahangi has been characterised by a number of biochemical and immunochemical means. The 30 kDa polypeptide is a glycoprotein which can be extracted from the worm surface by homogenization in the absence of detergents. The 30 kDa polypeptide can be metabolically labelled wit...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(88)90027-8
更新日期：1988-01-01 00:00:00
abstract：:gamma-Glutamyl transpeptidase, which is of central importance in the degradation of glutathione, was purified from Ascaris suum to apparent homogeneity. The enzyme was found to be a predominantly membrane-bound protein and was solubilized by Triton X-100. The purified enzyme, which exhibits a specific activity of 1009...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(96)02573-x
更新日期：1996-04-01 00:00:00
abstract：:Kinetoplast DNA (kDNA) was isolated from 56 stocks of Trypanosoma cruzi isolated from human patients, animals and insects from Brazil, Venezuela, Colombia and Costa Rica. Comparison of the patterns of digested kDNA on acrylamide gels led to the grouping of several stocks into two schizodemes. Schizodeme analysis was a...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(90)90160-n
更新日期：1990-09-01 00:00:00
abstract：:Amastigotes of Leishmania major were isolated from infected mice and radiolabeled for 2 h with [3H]galactose. An acidic [3H]glycoconjugate was extracted from a dilipidated residue fraction with the solvent water/ethanol/diethylether/pyridine/NH4OH (15:15:5:1:0.017). The radioactivity labeled glycoconjugate was found t...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(91)90030-a
更新日期：1991-03-01 00:00:00
abstract：:The azole antifungals ketoconazole and itraconazole possess in vitro antileishmanial activity against Leishmania mexicana mexicana amastigotes in macrophages (cell line J774G8). As in yeast and fungi, the activity is likely to be due to inhibition of the cytochrome P-450-dependent 14 alpha-demethylation of lanosterol ...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(89)90026-1
更新日期：1989-03-01 00:00:00
abstract：:Among several protease inhibitors tested, only leupeptin was found to modify qualitatively the processing of P126, a major antigen of the parasitophorous vacuole of Plasmodium falciparum, and to inhibit the release of merozoites. Whereas P126 is normally processed upon merozoite release into 2 polypeptides of 50 and 7...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(89)90029-7
更新日期：1989-03-01 00:00:00
abstract：:The alpha and beta tubulin genes of Leishmania major were cloned and used to study the genomic organization, chromosomal location and transcription of tubulin genes in L. major. The number of beta tubulin isogenes was determined by hybridization of probes representing the 5' and 3' ends of the cloned beta tubulin cDNA...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(87)90112-5
更新日期：1987-05-01 00:00:00
abstract：:Many surface antigens of the human malaria parasite Plasmodium falciparum show extraordinary diversity, with different alleles being so divergent as to be unalignable in some coding regions. To better understand the population history and modes of selection on such loci, we sequenced genomic regions flanking the highl...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2007.12.004
更新日期：2008-05-01 00:00:00
abstract：:The cyclophilins are a large family of proteins implicated in folding, transport and regulation of other proteins and are potential drug targets in cancer and in some viral and parasitic infections. The functionality of cyclophilins appears to depend on peptidyl-prolyl cis-trans isomerase (foldase) and/or molecular ch...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2012.04.006
更新日期：2012-07-01 00:00:00
abstract：:Giardia duodenalis is a widespread parasite of mammalian species, including humans. Due to its invariant morphology, investigation on aspects such as host specificity and transmission patterns requires a direct genetic characterization of cysts/trophozoites from host samples. A number of molecular assays have been dev...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章,评审
doi：10.1016/j.molbiopara.2008.04.006
更新日期：2008-08-01 00:00:00
abstract：:Intraerythrocytic Plasmodium falciparum digests vast amounts of hemoglobin within an acidic food vacuole (FV). Four homologous aspartic proteases participate in hemoglobin degradation within the FV. Plasmepsin (PM) I and II are thought to initiate degradation of the native hemoglobin molecule. PM IV and histo-aspartic...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(03)00119-1
更新日期：2003-07-01 00:00:00
abstract：:We have purified and characterized the Ascaris suum gamma-glutamylcysteine synthetase, the rate-limiting step in the glutathione biosynthesis. The purified enzyme exhibited a specific activity of 18 U (mg protein)-1. Estimation of the molecular mass of the native enzyme by FPLC on Superdex S-200 revealed the presence ...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(94)00064-t
更新日期：1995-06-01 00:00:00
abstract：:Infection with larval trematodes has been shown to inhibit several snail-host defences, including hemocyte phagocytosis, cytotoxicity, motility, and adherence. Certain plasma factors which mediate snail defence responses, and which may be produced by host hemocytes, also appear to be altered by these parasites. In thi...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(91)90124-o
更新日期：1991-11-01 00:00:00
abstract：:A myofibrillar protein extract has been isolated from the muscle of Ascaris suum. Two-dimensional electrophoresis of this extract revealed that the myosin light chain 1 (ALC1) migrates as 3 components with approximate isoelectric points in the range of 5.3-5.6. The most acidic component of ALC1 appeared to be phosphor...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(81)90007-4
更新日期：1981-06-01 00:00:00
abstract：:A restriction fragment from a cloned kinetoplast minicircle DNA has been shown to be diagnostic for Leishmania major. This 402-bp TaqI fragment has been used routinely (as a radiolabelled probe) to detect 10(4) parasites in simple dot blots, both experimentally and in epidemiological surveys. It positively identified ...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(89)90153-9
更新日期：1989-12-01 00:00:00
abstract：:Tubulin genes in Trypanosoma rangeli, the only trypanosome besides T. cruzi to infect humans in America, are organized in homogeneous, alternate alpha and beta gene tandem repeats of 3.8 kb. The basic repeat was cloned, mapped and partially sequenced. In contrast to most other eukaryotes, where tubulin genes are scatt...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(89)90054-6
更新日期：1989-05-15 00:00:00
abstract：:During intracellular development of the malarial parasite numerous membranous vesicles appear in the infected erythrocyte cytoplasm between the parasitophorous vacuolar membrane (PVM) and the erythrocyte plasma membrane. In this study we describe the characterization of a monoclonal antibody which recognizes two major...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(97)00206-5
更新日期：1998-03-15 00:00:00
abstract：:Studies measuring the glycolytic intermediate and adenine nucleotide concentrations in Trypanosoma brucei metabolising glucose either aerobically or under conditions where glycerol-3-phosphate oxidase is inactive have shown the following: 1. Inhibition with 0.5 mM salicylhydroxamic acid (SHAM) accurately simulates ana...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(80)90032-8
更新日期：1980-12-01 00:00:00
abstract：:We have successfully transfected Entamoeba histolytica trophozoites with constructs containing ferredoxin sequences fused to the reporter gene luciferase. We have determined the conditions and parameters necessary to maximise transient luciferase expression in our system. Our optimal construct gave values of 268 x 10(...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(95)02474-3
更新日期：1995-10-01 00:00:00
abstract：:Cyclic AMP-dependent protein kinase (protein kinase A, PKA) is a key element in many cell signaling pathways. An essential role of Plasmodium falciparum PKA (PfPKA) activity was reported in the intraerythrocytic growth of the malaria parasite. However, molecular characterization of PfPKA using purified recombinant pro...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2008.03.011
更新日期：2008-08-01 00:00:00
abstract：:After a single intravenous injection of suramin the rate of removal of the drug from the plasma into other tissue compartments of the rat is independent of initial concentration. The data can be fitted to the sum of two exponential functions, consistent with a two-compartment, open model system. Trypanosomes take up o...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(80)90050-x
更新日期：1980-10-01 00:00:00
abstract：:Both patent and prepatent adult Hymenolepis diminuta excreted 20-hydroxyecdysone into the culture medium when maintained in vitro. Patent worms also excreted ecdysone and comparatively large quantities of unidentified immunoreactive material of a relatively apolar nature. This latter material was shown to be depleted ...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(87)90075-2
更新日期：1987-12-01 00:00:00
