Purification and characterization of gamma-glutamylcysteine synthetase from Ascaris suum.


: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 of two enzyme activity peaks corresponding to molecular masses of 100 and 70 kDa. The higher-molecular-mass component could be dissociated by repeated gel filtration into the 70-kDa protein which is the enzymatically active subunit. The apparent Km values of the A. suum enzyme for L-aminobutyrate, L-cysteine and L-glutamate were 0.31, 0.41 and 0.94 mM, respectively. D,L-Buthionine-S,R-sulfoximine and cystamine showed time-dependent irreversible inhibitory effects on the A. suum enzyme activity with Ki values of 0.05 and 1.11 microM, respectively. The Ki values for the corresponding enzyme from rat kidney with D,L-buthionine-S,R-sulfoximine and cystamine were 7.19 and 22.2 microM, respectively. The time of half-inactivation of the enzyme at infinite concentration of D,L-buthionine-S,R-sulfoximine, tau 50, was determined to be 3.1 and 1.34 min, for the parasite and mammalian enzymes respectively. For cystamine, a tau 50 value of 3.32 min for the A. suum gamma-glutamylcysteine synthetase was determined, while a value of 2 min in case of rat kidney enzyme was found. The A. suum enzyme activity was competitively inhibited by glutathione with a Ki value of 0.11 mM.(ABSTRACT TRUNCATED AT 250 WORDS)


Mol Biochem Parasitol


Hussein AS,Walter RD




Has Abstract


1995-06-01 00:00:00














  • Characterisation of the gene encoding a 104-kilodalton microneme-rhoptry protein of Theileria parva.

    abstract::A neutralizing antiserum, C16, raised against sporozoites of Theileria parva parva was used to screen a lambda gt11 expression library of T. parva parva (Muguga) genomic DNA fragments. Proteins encoded by one phage clone, lambda TpS-17, were reactive with the C16 antiserum. Detailed characterisation of the DNA insert ...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Iams KP,Young JR,Nene V,Desai J,Webster P,ole-MoiYoi OK,Musoke AJ

    更新日期:1990-02-01 00:00:00

  • Stable transfection and continuous expression of heterologous genes in Entamoeba invadens.

    abstract::Amoebiasis is spread by the ingestion of dormant Entamoeba histolytica cysts. Intervention of encystation could break the transmission cycle, thereby reducing disease burden. The model system used to study trophozoite to cyst differentiation is Entamoeba invadens. Here we describe an electroporation-based method for s...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Singh N,Ojha S,Bhattacharya A,Bhattacharya S

    更新日期:2012-07-01 00:00:00

  • The secreted and somatic antigens of the third stage larva of Anisakis simplex, and antigenic relationship with Ascaris suum, Ascaris lumbricoides, and Toxocara canis.

    abstract::The in vitro-released 'excretory/secretory' (ES) and somatic antigens of the third stage (infective) larva of Anisakis simplex were characterised by radioiodination, immunoprecipitation, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Oral infection with the parasite elicited a substantial IgG antibody...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Kennedy MW,Tierney J,Ye P,McMonagle FA,McIntosh A,McLaughlin D,Smith JW

    更新日期:1988-10-01 00:00:00

  • Over-expression and localization of a host protein on the membrane of Cryptosporidium parvum infected epithelial cells.

    abstract::The genus Cryptosporidium includes several species of intestinal protozoan parasites which multiply in intestinal epithelial cells. The impact of this infection on the transcriptome of cultured host cells was investigated using DNA microarray hybridizations. The expression of 14 genes found to be consistently up- or d...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Yang YL,Serrano MG,Sheoran AS,Manque PA,Buck GA,Widmer G

    更新日期:2009-11-01 00:00:00

  • Sequencing and characterization of an Echinococcus multilocularis DNA probe and its use in the polymerase chain reaction.

    abstract::The nucleotide sequence of the cloned Echinococcus multilocularis DNA probe pAL1 was determined in order to simplify and improve the sensitivity of a diagnostic assay through the application of the polymerase chain reaction (PCR). The insert-specific oligonucleotides BG1 and BG2 define a 2.6-kb fragment in the genomic...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Gottstein B,Mowatt MR

    更新日期:1991-02-01 00:00:00

  • The Plasmodium falciparum exported protein PF3D7_0402000 binds to erythrocyte ankyrin and band 4.1.

    abstract::Plasmodium falciparum extensively modifies the infected red blood cell (RBC), resulting in changes in deformability, shape and surface properties. These alterations suggest that the RBC cytoskeleton is a major target for modification during infection. However, the molecular mechanisms leading to these changes are larg...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Shakya B,Penn WD,Nakayasu ES,LaCount DJ

    更新日期:2017-09-01 00:00:00

  • Biochemical characterization of Toxoplasma gondii 1-Cys peroxiredoxin 2 with mechanistic similarities to typical 2-Cys Prx.

    abstract::TgPrx2 represents a recently discovered cytosolic 1-Cys peroxiredoxin (Prx) from the intracellular parasite Toxoplasma gondii. Over-expression of the respective gene confers protection against H(2)O(2), suggesting that the protein possesses peroxidase activity. According to the current nomenclature eukaryotic typical ...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Deponte M,Becker K

    更新日期:2005-03-01 00:00:00

  • A gene-family encoding small exported proteins is conserved across Plasmodium genus.

    abstract::A gene-family, named sep, encoding small exported proteins conserved across Plasmodium species has been identified. SEP proteins (13-16 kDa) contain a predicted signal peptide at the NH(2)-terminus, an internal hydrophobic region and a polymorphic, low-complexity region at the carboxy-terminus. One member of the Plasm...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Birago C,Albanesi V,Silvestrini F,Picci L,Pizzi E,Alano P,Pace T,Ponzi M

    更新日期:2003-02-01 00:00:00

  • Molecular cloning and characterisation of a developmentally regulated putative metallopeptidase present in a host protective extract of Haemonchus contortus.

    abstract::Antisera from lambs immunised with the Haemonchus contortus integral membrane protein complex, Haemonchus galactose-containing glycoprotein (H-gal-GP), the lambs being refractory to subsequent challenge, were used to identify several clones from an adult H. contortus lambda gt11 cDNA library. Using gene-specific oligo...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Redmond DL,Knox DP,Newlands G,Smith WD

    更新日期:1997-03-01 00:00:00

  • SYBR, TaqMan, or both: highly sensitive, non-invasive detection of Cardicola blood fluke species in Southern Bluefin Tuna (Thunnus maccoyii).

    abstract::Three species of blood fluke from the genus Cardicola are known to parasitize and cause disease in Bluefin Tunas--C. forsteri, C. orientalis, and C. opisthorchis. Although initially believed to be separated by geography and host specificity, recent identification of at least two Cardicola spp. concurrently present wit...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Polinski M,Hamilton DB,Nowak B,Bridle A

    更新日期:2013-09-01 00:00:00

  • Identification of six Trypanosoma cruzi lineages by sequence-characterised amplified region markers.

    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


    authors: Brisse S,Dujardin JC,Tibayrenc M

    更新日期:2000-11-01 00:00:00

  • Insertion of the promoter for a variant surface glycoprotein gene expression site in an RNA polymerase II transcription unit of procyclic Trypanosoma brucei.

    abstract::The variant-specific surface glycoprotein (VSG) genes of Trypanosoma brucei are invariably expressed near the ends of chromosomes (telomeres). We have targeted a VSG gene expression site (ES) promoter driving a selectable marker gene (neomycin phosphotransferase) into a chromosome-internal transcription unit, the tubu...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Zomerdijk JC,Kieft R,Borst P

    更新日期:1993-02-01 00:00:00

  • Development of an untargeted metabolomics method for the analysis of human faecal samples using Cryptosporidium-infected samples.

    abstract::Faecal metabolite profiling, though in its infancy, allows for investigation of complex metabolic interactions between gastrointestinal infections or diseases and host health. In the present study, we describe a faecal metabolite extraction method for untargeted gas chromatography-mass spectrometry (GC-MS) analysis us...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Ng JS,Ryan U,Trengove RD,Maker GL

    更新日期:2012-10-01 00:00:00

  • Food vacuole plasmepsins are processed at a conserved site by an acidic convertase activity in Plasmodium falciparum.

    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


    authors: Banerjee R,Francis SE,Goldberg DE

    更新日期:2003-07-01 00:00:00

  • Pyrazolopyrimidine metabolism in African trypanosomes: metabolic similarities to Trypanosoma cruzi and Leishmania spp.

    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


    authors: Berens RL,Marr JJ,Brun R

    更新日期:1980-04-01 00:00:00

  • Surface epitope localization and gene structure of a Babesia bovis 44-kilodalton variable merozoite surface antigen.

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    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Jasmer DP,Reduker DW,Hines SA,Perryman LE,McGuire TC

    更新日期:1992-10-01 00:00:00

  • The influence of temperature and serum deprivation on the synthesis of heat-shock proteins and alpha and beta tubulin in promastigotes of Leishmania major.

    abstract::We have examined changes in the relative synthesis of individual proteins in promastigotes of Leishmania major subjected to decreasing serum levels in vitro. We observed increases in the relative synthesis of the putative heat-shock proteins of 82 and 70 kDa and of proteins of 79 and 41 kDa but decreases in the synthe...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Toye P,Remold H

    更新日期:1989-06-01 00:00:00

  • Crystal structure and putative substrate identification for the Entamoeba histolytica low molecular weight tyrosine phosphatase.

    abstract::Entamoeba histolytica is a eukaryotic intestinal parasite of humans, and is endemic in developing countries. We have characterized the E. histolytica putative low molecular weight protein tyrosine phosphatase (LMW-PTP). The structure for this amebic tyrosine phosphatase was solved, showing the ligand-induced conformat...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Linford AS,Jiang NM,Edwards TE,Sherman NE,Van Voorhis WC,Stewart LJ,Myler PJ,Staker BL,Petri WA Jr

    更新日期:2014-01-01 00:00:00

  • Characterization of macromolecular transport pathways in malaria-infected erythrocytes.

    abstract::We have previously provided evidence for a pathway in Plasmodium falciparum-infected erythrocytes, coined the parasitophorous duct pathway, which provides serum (macro)molecules direct access to intraerythrocytic parasites . The present study addresses the purity of the fluorescent macromolecules used to define the du...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Goodyer ID,Pouvelle B,Schneider TG,Trelka DP,Taraschi TF

    更新日期:1997-07-01 00:00:00

  • PI(3)P-independent and -dependent pathways function together in a vacuolar translocation sequence to target malarial proteins to the host erythrocyte.

    abstract::Malaria parasites export 'a secretome' of hundreds of proteins, including major virulence determinants, from their endoplasmic reticulum (ER), past the parasite plasma and vacuolar membranes to the host erythrocyte. The export mechanism is high affinity (nanomolar) binding of a host (cell) targeting (HT) motif RxLxE/D...

    journal_title:Molecular and biochemical parasitology

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    authors: Bhattacharjee S,Speicher KD,Stahelin RV,Speicher DW,Haldar K

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  • Expression profiling of the Leishmania life cycle: cDNA arrays identify developmentally regulated genes present but not annotated in the genome.

    abstract::As genomic sequencing of Leishmania nears completion, functional analyses that provide a global genetic perspective on biological processes are important. Despite polycistronic transcription, RNA transcript abundance can be measured using microarrays. To provide a resource to evaluate cDNA arrays, we undertook 5' expr...

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    authors: Almeida R,Gilmartin BJ,McCann SH,Norrish A,Ivens AC,Lawson D,Levick MP,Smith DF,Dyall SD,Vetrie D,Freeman TC,Coulson RM,Sampaio I,Schneider H,Blackwell JM

    更新日期:2004-07-01 00:00:00

  • High efficiency transfection of Plasmodium berghei facilitates novel selection procedures.

    abstract::The use of transfection in the study of the biology of malaria parasites has been limited due to poor transfection efficiencies (frequency of 10(-6) to 10(-9)) and a paucity of selection markers. Here, a new method of transfection, using non-viral Nucleofector technology, is described for the rodent parasite Plasmodiu...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Janse CJ,Franke-Fayard B,Mair GR,Ramesar J,Thiel C,Engelmann S,Matuschewski K,van Gemert GJ,Sauerwein RW,Waters AP

    更新日期:2006-01-01 00:00:00

  • Characterisation of the loci encoding the glutamic acid and alanine rich protein of Trypanosoma congolense.

    abstract::We have characterised the organisation of genes encoding the glutamate and alanine rich protein (GARP) surface coat of the procyclic and epimastigote stages of Trypanosoma congolense in the tsetse fly. The GARP genes are arranged at two, possibly physically linked, loci, one of which exhibits allelic variation. One lo...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Rangarajan D,Harvey TI,Barry JD

    更新日期:2000-02-05 00:00:00

  • The proteasome-ubiquitin pathway in the Schistosoma mansoni egg has development- and morphology-specific characteristics.

    abstract::Schistosoma mansoni eggs, consisting of an ovum surrounded by nutritive vitelline cells packaged in a tanned protein shell, are produced by paired worms residing in the mesenteric veins of the human host. The vitelline cells are degraded as the larval miracidium matures, the fully developed egg either crossing the gut...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Mathieson W,Castro-Borges W,Wilson RA

    更新日期:2011-02-01 00:00:00

  • mRNA sequences for Haemonchus contortus intestinal cathepsin B-like cysteine proteases display an extreme in abundance and diversity compared with other adult mammalian parasitic nematodes.

    abstract::Cathepsin B-like cysteine protease (cbl) genes produce the most abundant mRNAs ( approximately 16%) detected in the adult female intestine of the parasitic nematode Haemonchus contortus. CBL enzymes appear to digest host proteins and are vaccine candidates for immune control of H. contortus and potentially other paras...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Jasmer DP,Mitreva MD,McCarter JP

    更新日期:2004-10-01 00:00:00

  • Heterogeneity of the serine synthetic pathway in Entamoeba species.

    abstract::Phosphoserine phosphatase (PSP) catalyzes the third step of the phosphorylated serine biosynthetic pathway, and occurred multiple times in evolution, while enzymes catalyzing the first and second steps in the pathway have single respective origins. In the present study, we examined the existence of PSP among genus Ent...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Chiba Y,Makiuchi T,Jeelani G,Nozaki T

    更新日期:2016-06-01 00:00:00

  • Dominant lethality and insect population control.

    abstract::Recent advances in insect genetic engineering have opened up new possibilities in the genetic control of insect vectors of human diseases. We outline the current state of the sterile insect technique and show how the use of engineered dominant lethals can greatly increase the effectiveness of this approach. We conside...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章,评审


    authors: Alphey L,Andreasen M

    更新日期:2002-05-01 00:00:00

  • Cloning and sequencing of an hsp70 gene of Schistosoma mansoni.

    abstract::Schistosomes have a complex life cycle (vertebrate and molluscan hosts as well as larvae living freely in water) in which they are exposed to different environments and temperatures (20 degrees C - 37 degrees C). Since heat shock genes are activated in response to stress and during development [1], it is of interest t...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Neumann S,Ziv E,Lantner F,Schechter I

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  • Histone mediated gene activation in Toxoplasma gondii.

    abstract::DNA processes, including transcriptional activation, are largely governed by histone modifications. For most eukaryotes, chromatin-remodeling machines have been identified that move, replace, or decorate histones. Recent studies have shed some important light on similar machinery operating in apicomplexan parasites, i...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章,评审


    authors: Sullivan WJ Jr,Hakimi MA

    更新日期:2006-08-01 00:00:00

  • Polymorphism of proteins in malaria parasites following mefloquine treatment.

    abstract::Proteins in malaria parasites (Plasmodium falciparum) isolated from a patient in Thailand before treatment, and after recrudescence of infection subsequent to mefloquine treatment, were compared by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis. Nine 'pre-treatment' and six 'recrudescent' clones...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章


    authors: Pinswasdi C,Thaithong S,Beale GH,Fenton B,Webster HK,Pavanand K

    更新日期:1987-03-01 00:00:00