Activation of citrate synthase from a marine pseudomonad by adenosine monophosphate and potassium chloride

https://nsuworks.nova.edu/nsudigital_harrison/4028/thumbnail.jp

Structure, kinetic characterization and subcellular localization of the two ribulose 5-phosphate epimerase isoenzymes from Trypanosoma cruzi

The enzyme of the pentose phosphate pathway (PPP) ribulose-5-phosphate-epimerase (RPE) is encoded by two genes present in the genome of Trypanosoma cruzi CL Brener clone: TcRPE1 and TcRPE2. Despite high sequence similarity at the amino acid residue level, the recombinant isoenzymes show a strikingly different kinetics. Whereas TcRPE2 follows a typical michaelian behavior, TcRPE1 shows a complex kinetic pattern, displaying a biphasic curve, suggesting the coexistence of -at least-two kinetically different molecular forms. Regarding the subcellular localization in epimastigotes, whereas TcRPE1 is a cytosolic enzyme, TcRPE2 is localized in glycosomes. To our knowledge, TcRPE2 is the first PPP isoenzyme that is exclusively localized in glycosomes. Over-expression of TcRPE1, but not of TcRPE2, significantly reduces the parasite doubling time in vitro, as compared with wild type epimastigotes. Both TcRPEs represent single domain proteins exhibiting the classical α/β TIM-barrel fold, as expected for enzymes with this activity. With regard to the architecture of the active site, all the important amino acid residues for catalysis -with the exception of M58- are also present in both TcRPEs models. The superimposition of the binding pocket of both isoenzyme models shows that they adopt essentially identical positions in the active site with a residue specific RMSD < 2Å, with the sole exception of S12, which displays a large deviation (residue specific RMSD: 11.07 A). Studies on the quaternary arrangement of these isoenzymes reveal that both are present in a mixture of various oligomeric species made up of an even number of molecules, probably pointing to the dimer as their minimal functional unit. This multiplicity of oligomeric species has not been reported for any of the other RPEs studied so far and it might bear implications for the regulation of TcRPEs activity, although further investigation will be necessary to unravel the physiological significance of these structural findings.Fil: Gonzalez, Soledad Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); ArgentinaFil: Valsecchi, Wanda Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Maugeri, Dante. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); ArgentinaFil: Delfino, Jose Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Cazzulo, Juan Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentin

An accurate record of volcanic ash fall deposition as characterized by dispersed organic matter in a Lower Permian tonstein bed (Faxinal Coalfield, Paraná Basin, Brazil)

For the first time, the dispersed organic matter in the tonstein layer interbedded with a coal seam in the Faxinal Coalfield (Sakmarian, Southern Paraná Basin, Brazil) is characterized. The deposition of clusters of pollen grains was highly influenced by the intense ash fall process that probably occurred during seasonal dehiscence of reproductive structures. The well-preserved phytoclasts with their upper and lower leaf cuticles stuck together indicate that the rapid fall of ash on this material hindered organic biodegradation. The preservation of seemingly autochthonous Botryococcus colonies at the top of the tonstein layer is evidence of the subaqueous deposition of this layer. The darkening in cuticles and xylem phytoclasts can be attributed to different causes: the thermal influence of ash fall during deposition, chemical effects of the ash, prolonged oxidation of organic matter in low water level conditions or the burning of plant organs by wildfires. Analyses of dispersed organic matter along the tonstein layer showed that the organic matter succession reflects the composition of different plant strata (herbaceous pteridophytes and arboreal glossopterids-cordaitaleans) around the deposition site

Tetrameric and dimeric malate dehydrogenase isoenzymes in Trypanosoma cruzi epimastigotes

Two malate dehydrogenase isoforms, named MDH1 and MDH2, have been purified to homogeneity from Trypanosoma cruzi epimastigotes. Both enzymes consist of subunits with a molecular mass close to 33 kDa; native molecular mass determination by gel filtration, however, indicated that MDH1 is a dimer, whereas MDH2 is a tetramer. Both isoforms did not cross-react immunologically. The N-termini of both MDH isoforms and several tryptic peptides of MDH1 (amounting to about one third of the complete molecule) have been sequenced by automated Edman degradation. The tryptic digests of both enzymes have also been analysed by mass spectrometry (MALDI-TOF MS). The apparent K(m) values in both directions of the reaction have been determined, as well as the possible inhibition by excess of the substrate oxaloacetate. The sequence data, together with the pI values and the presence or absence of oxaloacetate inhibition indicate that the dimeric MDH1 is the mitochondrial isoenzyme, whereas the tetrameric MDH2 is the glycosomal isoenzyme. No evidence was found for the presence of a cytosolic isoform. (C) 2000 Elsevier Science B.V.Fil: Reynoso Hunter, Giselle. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Hellman, Ulf. Ludwig Institute for Cancer Research; SueciaFil: Cazzulo, Juan Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Nowicki, Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentin

The C-terminal region of Trypanosoma cruzi MASPs is antigenic and secreted via exovesicles.

Trypanosoma cruzi is the etiological agent of Chagas disease, a neglected and emerging tropical disease, endemic to South America and present in non-endemic regions due to human migration. The MASP multigene family is specific to T. cruzi, accounting for 6% of the parasite's genome and plays a key role in immune evasion. A common feature of MASPs is the presence of two conserved regions: an N-terminal region codifying for signal peptide and a C-terminal (C-term) region, which potentially acts as GPI-addition signal peptide. Our aim was the analysis of the presence of an immune response against the MASP C-term region. We found that this region is highly conserved, released via exovesicles (EVs) and has an associated immune response as revealed by epitope affinity mapping, IFA and inhibition of the complement lysis assays. We also demonstrate the presence of a fast IgM response in Balb/c mice infected with T. cruzi. Our results reveal the presence of non-canonical secreted peptides in EVs, which can subsequently be exposed to the immune system with a potential role in evading immune system targets in the parasite

Chagas' disease: an update on immune mechanisms and therapeutic strategies

The final decade of the 20th century was marked by an alarming resurgence in infectious diseases caused by tropical parasites belonging to the kinetoplastid protozoan order. Among the pathogenic trypanosomatids, some species are of particular interest due to their medical importance. These species include the agent responsible for Chagas' disease, Trypanosoma cruzi. Approximately 8 to 10 million people are infected in the Americas, and approximately 40 million are at risk. in the present review, we discuss in detail the immune mechanisms elicited during infection by T. cruzi and the effects of chemotherapy in controlling parasite proliferation and on the host immune system.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Instituto Nacional de Biologia Estrutural e QuImica Medicinal em Doencas Infecciosas (INBEQMeDI)Univ São Paulo, Inst Biomed Sci, Dept Parasitol, São Paulo, BrazilMackenzie Presbeterian Univ, Ctr Biomol Sci & Hlth, São Paulo, BrazilNatl Univ Rosario, Sch Med Sci, Inst Immunol, Rosario, Santa Fe, ArgentinaCSIC, Inst Parasitol & Biomed Lopez Neyra, Granada, SpainUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilFAPESP: 08/57596-4FAPESP: 07/08648-9CNPq: 473906/2008-2Web of Scienc

Análise Preliminar de Cutículas Dispersas e Outros Microfragmentos Vegetais Encontrados em Carvões da Camada Morungava 6 e Folhelhos Carbonosos, Bacia Carbonífera de Gravataí, R.G.S. (Bacia do Paraná)

São analisados neste trabalho microfragmentos vegetais encontrados em associação com material palinológico resgatado em amostras de carvão da Camada Morungava 6 e folhelhos carbonosos (Bacia Carbonífera de Gravataí, Rio Grande do Sul), incluindo cutículas dispersas e restos de tecido xilemático, parenquimático e esclerenquimático. A análise quantitativa do material, feita de modo preliminar, revelou a predominância de cutículas relacionáveis a folhas e frutificações de Glossopteridales, Cordaitales e Sphenophyllales em comparação a fragmentos de tecidos vinculáveis a córtex e xilema. A classificação utilizada não teve por base um sistema taxonômico definido mas abrange, parcialmente, critérios sistemáticos adotados por MEYEN (1965), BOSE & MAHESHWARI (1968), CHANDRA & SURANGE (1975), NAVALE (1976) e MAITHY (1976)

An accurate record of volcanic ash fall deposition as characterized by dispersed organic matter in a lower Permian tonstein layer (Faxinal Coalfield, Paraná Basin, Brazil)

For the first time, the dispersed organic matter in the tonstein layer interbedded with a coal seam in the Faxinal Coalfield (Sakmarian, Southern Paraná Basin, Brazil) is characterized. The deposition of clusters of pollen grains was highly influenced by the intense ash fall process that probably occurred during seasonal dehiscence of reproductive structures. The well-preserved phytoclasts with their upper and lower leaf cuticles stuck together indicate that the rapid fall of ash on this material hindered organic biodegradation. The preservation of seemingly autochthonous Botryococcus colonies at the top of the tonstein layer is evidence of the subaqueous deposition of this layer. The darkening in cuticles and xylem phytoclasts can be attributed to different causes: the thermal influence of ash fall during deposition, chemical effects of the ash, prolonged oxidation of organic matter in low water level conditions or the burning of plant organs by wildfires. Analyses of dispersed organic matter along the tonstein layer showed that the organic matter succession reflects the composition of different plant strata (herbaceous pteridophytes and arboreal glossopterids-cordaitaleans) around the deposition site