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Improving the study of proton transfers between amino acid sidechains in solution: choosing appropriate DFT functionals and avoiding hidden pitfalls

Silva, Pedro J.; Perez, Marta A. S.; Brás, Natércia F.; Fernandes, Pedro A.; Ramos, Maria João
Fonte: Springer Publicador: Springer
Tipo: Artigo de Revista Científica
Publicado em //2012 Português
Relevância na Pesquisa
65.99%
We have studied the influence of implicit solvent models, inclusion of explicit water molecules, inclusion of vibrational effects, and density functionals on the quality of the predicted pK a of small amino acid side chain models. We found that the inclusion of vibrational effects and explicit water molecules is crucial to improve the correlation between the computed and the experimental values. In these micro-solvated systems, the best agreement between DFT-computed electronic energies and benchmark values is afforded by BHHLYP and B97-2. However, approaching experimental results requires the addition of more than three explicit water molecules, which generates new problems related to the presence of multiple minima in the potential energy surface. It thus appears that a satisfactory ab initio prediction of amino acid side chain pK a will require methods that sample the configurational space in the presence of large solvation shells, while at the same time computing vibrational contributions to the enthalpy and entropy of the system under study in all points of that surface. Pending development of efficient algorithms for those computations, we strongly suggest that whenever counterintuitive protonation states are found in a computational study (e.g....

Antiviral and antiparasite properties of an L-amino acid oxidase from the Snake Bothrops jararaca: Cloning and identification of a complete cDNA sequence (Retracted article. See vol. 80, pg. 288, 2010)

SANT`ANA, Carolina D.; MENALDO, Danilo L.; COSTA, Tassia R.; GODOY, Harryson; MULLER, Vanessa D. M.; AQUINO, Victor H.; ALBUQUERQUE, Sergio; SAMPAIO, Suely V.; MONTEIRO, Marta C.; STABELI, Rodrigo G.; SOARES, Andreimar M.
Fonte: PERGAMON-ELSEVIER SCIENCE LTD Publicador: PERGAMON-ELSEVIER SCIENCE LTD
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
66.07%
L-Amino acid oxidases (LAAOs, EC 1.4.3.2) are flavoenzymes that catalyze the stereospecific oxidative deamination of an L-amino acid substrate to the corresponding a-ketoacid with hydrogen peroxide and ammonia production. The present work describes the first report on the antiviral (Dengue virus) and antiprotozoal (trypanocidal and leishmanicide) activities of a Bothrops jararaca L-amino acid oxidase (BjarLAAO-I) and identify its cDNA sequence. Antiparasite effects were inhibited by catalase, suggesting that they are mediated by H(2)O(2) production. Cells infected with DENV-3 virus previously treated with BjarLAAO-I, showed a decrease in viral titer (13-83-fold) when compared with cells infected with untreated viruses. Untreated and treated promastigotes (T. cruzi and L. amazonensis) were observed by transmission electron microscopy with different degrees of damage. Its complete cDNA sequence, with 1452 bp, encoded an open reading frame of 484 amino acid residues with a theoretical molecular weight and pl of 54,771.8 and 5.7, respectively. The cDNA-deduced amino acid sequence of BjarLAAO shows high identity to LAAOs from other snake venoms. Further investigations will be focused on the related molecular and functional correlation of these enzymes. Such a study should provide valuable information for the therapeutic development of new generations of microbicidal drugs. (C) 2008 Elsevier Inc. All rights reserved.

Structural and functional properties of Bp-LAAO, a new L-amino acid oxidase isolated from Bothrops pauloensis snake venom

RODRIGUES, Renata S.; SILVA, Juliana F. da; FRANCA, Joharal Boldrini; FONSECA, Fernando P. P.; OTAVIANO, Antonio R.; SILVA, Flavio Henrique; HAMAGUCHI, Amelia; MAGRO, Angelo J.; BRAZ, Antonio Sergio K.; SANTOS, Juliana I. dos; HOMSI-BRANDEBURGO, Maria Ine
Fonte: ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER Publicador: ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
65.97%
An L-amino acid oxidase (Bp-LAAO) from Bothrops pauloensis snake venom was highly purified using sequential chromatography steps on CM-Sepharose, Phenyl-Sepharose CL4B, Benzamidine Sepharose and C18 reverse-phase HPLC. Purified Bp-LAAO showed to be a homodimeric acidic glycoprotein with molecular weight around 65 kDa under reducing conditions in SDS-PAGE. The best substrates for Bp-LAAO were L-Met, L-Leu, L-Phe and L-Ile and the enzyme showed a strong reduction of its catalytic activity upon L-Met and L-Phe substrates at extreme temperatures. Bp-LAAO showed leishmanicidal, antitumoral and bactericidal activities dose dependently. Bp-LAAO induced platelet aggregation in platelet-rich plasma and this activity was inhibited by catalase. Bp-LAAC-cDNA of 1548 bp codified a mature protein with 516 amino acid residues corresponding to a theoretical isoelectric point and molecular weight of 6.3 and 58 kDa, respectively. Additionally, structural and phylogenetic studies identified residues under positive selection and their probable location in Elp-LAAO and other snake venom LAAOs (svLAAOs). Structural and functional investigations of these enzymes can contribute to the advancement of toxinology and to the elaboration of novel therapeutic agents. (C) 2009 Elsevier Masson SAS. All rights reserved.; Fundacao de Amparo a Pesquisa de Minas Gerais (FAPEMIG); Sao Paulo (FAPESP); Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)...

Lipid, fatty acid, protein, amino acid and ash contents in four Brazilian red algae species

GRESSLER, Vanessa; YOKOYA, Nair Sumie; FUJII, Mutue Toyota; COLEPICOLO, Pio; MANCINI FILHO, Jorge; TORRES, Rosangela Pavan; PINTO, Ernani
Fonte: ELSEVIER SCI LTD Publicador: ELSEVIER SCI LTD
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
66.05%
Four species of marine benthic algae (Laurencia filiformis, L. intricata, Gracilaria domingensis and G. birdiae) that belong to the phylum Rhodophyta were collected in Espirito Santo State, Brazil and investigated concerning their biochemical composition (fatty acid, total lipid, soluble proteins, amino acid and ash). The total content of lipid (% dry weight) ranged from 1.1% to 6.2%: fatty acid from 0.7% to 1.0%: soluble protein from 4.6% to 18.3%, amino acid from 6.7% to 11.3% and ash from 22.5% to 38.4%. judging from their composition, the four species of algae appear to be potential sources of dietary proteins, amino acids, lipids and essential fatty acids for humans and animals. (C) 2009 Elsevier Ltd. All rights reserved.; FAPESP (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo); Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); CAPES (Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico); Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Isolamento e caracterização bioquímica e funcional de L-aminoácido oxidase do veneno de 'Bothrops atrox'; Isolation and characterization of an L-amino acid oxidase from Bothrops atrox snake venom

Alves, Raquel de Melo
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: dissertação de mestrado Formato: application/pdf
Publicado em 16/03/2007 Português
Relevância na Pesquisa
66.07%
Os venenos de serpentes são ricos em proteínas, enzimas e peptídeos biologicamente ativos. Diversas enzimas tais como fosfolipases A2 (PLA2s), metaloproteases (MP), L-aminoácido oxidases (LAAOs) são responsáveis pelo quadro clínico do envenenamento ofídico. Atualmente, o isolamento e a caracterização funcional e estrutural destas enzimas têm auxiliado na compreensão do mecanismo de ação destas toxinas e nas formas alternativas de tratamento. Além disso, estas enzimas se tornaram valiosas ferramentas de aplicação biotecnológica, na busca de novos fármacos de interesse na clínica médica. O objetivo deste trabalho foi isolar e caracterizar bioquímica e funcionalmente a L-aminoácido oxidase do veneno de Bothrops atrox. O isolamento da LAAO de B. atrox (LAAOBatrox) envolveu três etapas cromatográficas: a exclusão molecular em Sephadex G-75, troca iônica em ES-502N utilizando CLAE e, por último, a cromatografia de afinidade com uso da Lentil-Lectina, conferindo um alto grau de pureza à proteína confirmado por SDS-PAGE. Após a caracterização bioquímica, a LAAOBatrox demonstrou ser uma glicoproteína com 12% de açúcar, massa molecular relativa de 67.000 e pI de 4,4 confirmando seu caráter ácido pela composição em aminoácidos...

Caracterização bioquímica, estrutural e funcional de uma L-aminoácido oxidase isolada de peçonha de Lachesis muta (Serpentes, Viperidae); Purification, biochemistry and functional characterization of a new L-amino acid oxidase from Lachesis muta venom.

Silva, Cristiane Bregge da
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Tese de Doutorado Formato: application/pdf
Publicado em 31/10/2011 Português
Relevância na Pesquisa
66.13%
As peçonhas de serpentes contêm uma mistura complexa de substâncias farmacologicamente ativas, como metaloproteases, fosfolipases A2, serino-proteases, L-aminoácido oxidase (LAAO), além de outros importantes compostos sem ação enzimática. LAAOs são flavoproteínas que catalisam a desaminação oxidativa de L-aminoácidos e produzem o -cetoácido correspondente, com a concomitante liberação de amônia e peróxido de hidrogênio. A peçonha de Lachesis muta (L. muta) contém L-aminoácido oxidase, a qual pode contribuir com o envenenamento. Portanto, o objetivo deste trabalho é a purificação da L-amino acido oxidase de peçonha de Lachesis muta (LmLAAO) e a sua caracterização bioquímica, estrutural e funcional. Para isso, foram desenvolvidos dois protocolos distintos de purificação, os quais forneceram LmLAAO com grande pureza. No primeiro protocolo, 20 mg de peçonha bruta de L. muta foram submetidos a uma gel filtração em Sephacryl S100®. Das dez frações obtidas, a primeira fração apresentou atividade L-aminoácido oxidase e foi submetida a mais um passo cromatográfico em Mono Q®. A homogeneidade da fração com atividade L-aminoácido oxidase após a troca iônica foi comprovada por presença de banda única com 60...

Caracterização funcional e estudo dos mecanismos de resposta ao dano celular causado por uma L-aminoácido oxidase de Calloselasma rhodostoma em linhagens celulares humanas; Functional characterization and study of the mechanisms of response to the cellular damage caused by an L-amino acid oxidase from Calloselasma rhodostoma in human cell lines

Costa, Tássia Rafaella
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Tese de Doutorado Formato: application/pdf
Publicado em 05/12/2014 Português
Relevância na Pesquisa
65.97%
As L-aminoácido oxidases (LAAOs) isoladas de peçonhas de serpentes são alvos de um grande número de pesquisas devido às suas inúmeras ações biológicas e farmacológicas. O objetivo do presente trabalho foi caracterizar funcionalmente a Laminoácido oxidase da peçonha de Calloselasma rhodostoma (CR-LAAO) por meio das atividades: bactericida, fungicida, leishmanicida, tripanomicida, citotóxica, inflamatória e análise da expressão de genes e proteínas apoptóticas. A CR-LAAO mostrou-se altamente citotóxica sobre as células tumorais HepG2 e HL-60, promovendo cerca de 80% de morte celular na maior concentração testada (100 ?g/mL) e apresentou baixa toxicidade sobre células PBMC. Foi possível observar que a proteína induziu apoptose (AV+) em PBMC. Em HepG2, as menores concentrações (0,1-2,5 ?g/mL) causaram apoptose (AV+), e as maiores (5-100 ?g/mL) causaram apoptose/necrose (PI+/AV+). Em HL-60, as concentrações testadas (0,1-100 ?g/mL) induziram apoptose/necrose (PI+/AV+). A expressão do gene FAS e ativação das caspases 8 e 3 determinou a ativação da via extrínseca da apoptose na linhagem HL-60. A CR-LAAO promoveu algumas alterações na modulação do ciclo celular, sendo que nas linhagens tumorais os atrasos se concentraram nas fases G0/G1 e S do ciclo celular. Ademais...

Effect of energy levels of diets formulated on total or digestible amino acid basis on broiler performance

Maiorka, Alex; Dahlke, Fabiano; Santin, E.; Kessler, Alexandre de Mello; Penz Junior, Antonio Mario
Tipo: Artigo de Revista Científica Formato: application/pdf
Português
Relevância na Pesquisa
65.97%
This study was conducted to evaluate the performance and carcass and breast yields of broilers fed two different energy levels (3,200 or 2,900 kcal ME/kg) and two ways of expressing amino acid requirements (total -TAA or digestible - DAA) between 21 and 42 days of age. The results showed that broilers fed the diet containing 3,200 kcal ME/kg had higher weight gain (P=0.015), better feed conversion (P=0.001), and higher abdominal fat deposition (P=0.001) as compared to those fed the diet containing 2,900 kcal ME/kg. Diets formulated on DAA basis promoted higher weight gain (P=0.043), better feed conversion (P=0.010) and better conversion of ME intake into weight gain (P=0.007) as compared to those formulated on TAA basis. The results of this study suggest that formulation based on DAA is necessary if the diets contain protein sources that are not reliable in terms of amino acid digestibility. The response to formulation based on DAA was minimized when birds received the low energy level diet (2,900 kcal ME/kg).

Antibacterial and antiparasitic effects of Bothrops marajoensis venom and its fractions: Phospholipase A(2) and L-amino acid oxidase

Costa Torres, Alba Fabiola; Dantas, Rodrigo Tavares; Toyama, Marcos H.; Diz Filho, Eduardo; Zara, Fernando Jose; Rodrigues de Queiroz, Maria Goretti; Pinto Nogueira, Nadia Accioly; de Oliveira, Marcia Rosa; Toyama, Daniela de Oliveira; Monteiro, Helena S.
Fonte: Pergamon-Elsevier B.V. Ltd Publicador: Pergamon-Elsevier B.V. Ltd
Tipo: Artigo de Revista Científica Formato: 795-804
Português
Relevância na Pesquisa
66.09%
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq); Some proteins present in snake venom possess enzymatic activities, such as phospholipase A(2) and L-amino acid oxidase. In this study, we verify the action of the Bothrops marajoensis venom (BmarTV), PLA(2) (BmarPLA(2)) and LAAO (BmarLAAO) on strains of bacteria, yeast, and Leishmania sp. The BmarTV was isolated by Protein Pack 5PW, and several fractions were obtained. Reverse phase HPLC showed that BmarPLA(2) was isolated from the venom, and N-terminal amino acid sequencing of sPLA(2) showed high amino acid identity with other lysine K49 sPLA(2)s isolated from Bothrops snakes. The BmarLAAO was purified to high molecular homogeneity and its N-terminal amino acid sequence demonstrated a high degree of amino acid conservation with others LAAOs. BmarLAAO was able to inhibit the growth of P. aeruginosa, C. albicans and S. aureus in a dose-dependent manner. The inhibitory effect was more significant on S. aureus, with a MIC = 50 mu g/mL and MLC = 200 mu g/mL However, the BmarTV and BmarPLA(2) did not demonstrate inhibitory capacity. BmarLAAO was able to inhibit the growth of promastigote forms of L chagasi and L amazonensis, with an IC50 = 2.55 mu g/mL and 2.86 mu g/mL for L. amazonensis and L. chagasi...

Pearl millet utilization in ccommercial laying hen diets formulated on a total or digestible amino acid basis

Filardi, RS; Junqueira, Otto Mack; Casartelli, em; Laurentiz, AC; Duarte, KF; Assuena, V
Fonte: Fundação APINCO de Ciência e Tecnologia Avícolas Publicador: Fundação APINCO de Ciência e Tecnologia Avícolas
Tipo: Artigo de Revista Científica Formato: 99-105
Português
Relevância na Pesquisa
66.01%
An experiment was carried out to evaluate the effect of replacing corn with pearl millet in commercial layer diets, formulated according to the minimal requirements for total and digestible amino acids. Two hundred and forty Lohmann LSL laying hens with 25 weeks of age were distributed in a completely randomized experimental design according to a 2 x 5 factorial arrangement with 3 replicates of 8 birds. Feed was formulated on two amino acid basis (total or digestible) according to Rostagno et al. (2000) and there were five pearl millet inclusion levels (0%, 25%, 50%, 75%, and 100%). Performance and egg quality were evaluated during five periods of 21 days.At the end of each period, feed intake, egg production, egg weight and feed conversion were evaluated. In the last three days of each period, the following egg quality parameters were evaluated: Haugh Unit, yolk pigmentation index, egg specific weight, shell percentage and shell thickness. Digestible amino acid requirements resulted in decreased feed intake (p

Amino acid, Antioxidant and Ion Profiles of Carpolobia lutea Leaf (Polygalaceae)

Nwidu, Lucky Legbosi; Cilli, Eduardo Maffud; Vilegas, Wagner
Fonte: Pharmacotherapy Group Publicador: Pharmacotherapy Group
Tipo: Artigo de Revista Científica Formato: 807-813
Português
Relevância na Pesquisa
66.03%
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); Purpose: To evaluate the amino acid, antioxidant and ionic profiles of Carpolobia lutea leaf (Polygalaceae) extract (CLL).Methods: The powdered leaf was macerated and subjected to gradient solvent extraction with n-hexane, chloroform, ethyl acetate and ethanol for 72 h to obtain their respective fractions. Amino acid analysis was by cation-exchange chromatography using automated amino acid analyser. Antioxidant potential was obtained by spectrophotometric assay using 2, 2-Diphenyl-1-picrylhydrazyl DPPH while elemental and ionic analyses were carried out by atomic absorption spectrophotometry and potentiometric titration, respectively.Results: Proline, alanine, serine, valine, glycine, glutamate and lysine were found in the ethanol fraction while lysine, phenyl alanine, glycine and serine were present in the ethyl acetate fraction but not in the non-polar fractions, n-hexane and chloroform. The ethyl acetate fraction contained more lysine, phenyl alanine, glycine and serine the other leaf fractions. Minimal radical scavenging activity of all the fractions was recorded. The most abundant cations in the extract were potassium and phosphorus (2.16 +/- 0.05 and 1.90 +/- 0.06 mg/g...

Eucalyptus ESTs associated with resistance to herbicide inhibitors of aromatic and branched-chain amino acid synthesis

Velini, Edivaldo Domingues; Trindade, Maria Lúcia Bueno; Alves, Elza; Catâneo, Ana Catarina; Marino, Celso Luis; de Godoy Maia, Ivan; Mori, Edson Seizo; Furtado, Edson Luiz; Guerrini, Iraê Amaral; Wilcken, Carlos Frederico
Tipo: Artigo de Revista Científica Formato: 575-581
Português
Relevância na Pesquisa
66.05%
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); Herbicides inhibit enzymatic systems of plants. Acetolactate synthase (ALS, EC = 4.1.3.18) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, EC 2.5.1.19) are key enzymes for herbicide action. Hundreds of compounds inhibit ALS. This enzyme is highly variable, enabling the selective control of weeds in a number of crops. Glyphosate, the only commercial herbicide inhibiting EPSPS is widely used for non-selective control of weeds in many crops. Recently, transgenic crops resistant to glyphosate were developed and have been used by farmers. The aim of this study was the data mining of eucalypt expressed sequence tags (ESTs) in the FORESTs Genome Project database (https://forests.esalq.usp.br) related to these enzymes. Representative amino acid sequences from the NCBI database associated with ALS and EPSPS were blasted with ESTs from the FORESTs database using the tBLASTx option of the blast tool. The best blasting reads and clusters from FORESTs, represented as nucleotide sequences, were blasted back with the NCBI database to evaluate the level of similarity with available sequences from different species. One and seven clusters were identified as showing high similarity with EPSPS and ALS sequences from the literature...

Structural insights into selectivity and cofactor binding in snake venom l-amino acid oxidases

Ullah, A.; Souza, T. A C B; Abrego, J. R B; Betzel, C.; Murakami, M. T.; Arni, R. K.
Tipo: Artigo de Revista Científica Formato: 124-128
Português
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66.09%
l-Amino acid oxidases (LAAOs) are flavoenzymes that catalytically deaminate l-amino acids to corresponding ?-keto acids with the concomitant production of ammonia (NH 3) and hydrogen peroxide (H 2O 2). Particularly, snake venom LAAOs have been attracted much attention due to their diverse clinical and biological effects, interfering on human coagulation factors and being cytotoxic against some pathogenic bacteria and Leishmania ssp. In this work, a new LAAO from Bothrops jararacussu venom (BjsuLAAO) was purified, functionally characterized and its structure determined by X-ray crystallography at 3.1å resolution. BjsuLAAO showed high catalytic specificity for aromatic and aliphatic large side-chain amino acids. Comparative structural analysis with prokaryotic LAAOs, which exhibit low specificity, indicates the importance of the active-site volume in modulating enzyme selectivity. Surprisingly, the flavin adenine dinucleotide (FAD) cofactor was found in a different orientation canonically described for both prokaryotic and eukaryotic LAAOs. In this new conformational state, the adenosyl group is flipped towards the 62-71 loop, being stabilized by several hydrogen-bond interactions, which is equally stable to the classical binding mode. © 2012 Elsevier Inc.

Soy protein concentrate as a protein source for Senegalese sole (Solea senegalensis Kaup 1858) diets: Effects on growth and amino acid metabolism of postlarvae

Aragão, C.; Conceição, L. E. C.; Dias, J.; Marques, A. C.; Gomes, E.; Dinis, Maria Teresa
Fonte: Wiley Publicador: Wiley
Tipo: Artigo de Revista Científica
Publicado em //2003 Português
Relevância na Pesquisa
66.06%
The objective of this workwas to evaluate the effect of a dietary amino acid imbalance, originating from the use of a soy protein concentrate (SPC) as the major protein source, on the growth performance and amino acid metabolism of Senegalese sole (Solea senegalensis) postlarvae. Senegalese sole (85.6724.6mg wet weight) were fed one of two experimental diets: one based on fish meal (FM) and another based on SPC. Diet were isonitrogenous (around 56% crude protein) and isoenergetic. Diet acceptability was very good and the growth rate was 6.9%day 1 for sole eating the FM diet and 6.0%day 1 for sole eating the SPC diet. Mass-speci¢c ammonia excretion and the activities of selected amino acid metabolic enzymes (ALAT, ASATand GDH) did not present significant differences between treatments, although this may have been due to the high variability found for these parameters in the SPC treatment. This variability may suggest different capacities of individual fish to adapt to the possible methionine dietary deficiency. The utilization of amino acids as a substrate for lipogenesis does not seem to be affected by the dietary protein source, since NAPDH-generating enzymes (G6PD and ME) had similar activities in both treatments. Amino acid metabolism in Senegalese sole postlarvae seems to be slightly affected by the dietary protein source. Nevertheless...

Amino acid composition of parturient plasma, the intervillous space of the placenta and the umbilical vein of term newborn infants

Camelo Jr.,J.S.; Jorge,S.M.; Martinez,F.E.
Fonte: Associação Brasileira de Divulgação Científica Publicador: Associação Brasileira de Divulgação Científica
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/05/2004 Português
Relevância na Pesquisa
66.12%
The objective of the present study was to determine the levels of amino acids in maternal plasma, placental intervillous space and fetal umbilical vein in order to identify the similarities and differences in amino acid levels in these compartments of 15 term newborns from normal pregnancies and deliveries. All amino acids, except tryptophan, were present in at least 186% higher concentrations in the intervillous space than in maternal venous blood, with the difference being statistically significant. This result contradicted the initial hypothesis of the study that the plasma amino acid levels in the placental intervillous space should be similar to those of maternal plasma. When the maternal venous compartment was compared with the umbilical vein, we observed values 103% higher on the fetal side which is compatible with currently accepted mechanisms of active amino acid transport. Amino acid levels of the placental intervillous space were similar to the values of the umbilical vein except for proline, glycine and aspartic acid, whose levels were significantly higher than fetal umbilical vein levels (average 107% higher). The elevated levels of the intervillous space are compatible with syncytiotrophoblast activity, which maintain high concentrations of free amino acids inside syncytiotrophoblast cells...

Amniotic fluid amino acid levels in non-immune hydrops fetalis: a case-control study

Erdemo?lu,M.; Kuyumcuoglu,U.; Guzel,A.I.; Celik,Y.; Kale,E.
Fonte: Associação Brasileira de Divulgação Científica Publicador: Associação Brasileira de Divulgação Científica
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/07/2011 Português
Relevância na Pesquisa
66.07%
In a prospective case-control study, we compared the amniotic fluid amino acid levels in non-immune hydrops fetalis (NIHF) and normal fetuses. Eighty fetuses underwent amniocentesis for different reasons at the prenatal diagnosis unit of the Department of Obstetrics and Gynecology, Faculty of Medicine, Dicle University. Forty of these fetuses were diagnosed with NIHF. The study included 40 women each in the NIHF (mean age: 27.69 ± 4.56 years) and control (27.52 ± 5.49 years) groups, who had abnormal double- or triple-screening test values with normal fetuses with gestational ages of 23.26 ± 1.98 and 23.68 ± 1.49 weeks at the time of sample collection, respectively. Amniotic fluid amino acid concentrations (intra-assay variation: 2.26-7.85%; interassay variation: 3.45-8.22%) were measured using EZ:faast kits (EZ:faast GC/FID free (physiological) amino acid kit; Phenomenex, USA) by gas chromatography. The standard for quantitation was a mixture of free amino acids from Phenomenex. The levels of 21 amino acids were measured. The mean phosphoserine and serine levels were significantly lower in the NIHF group, while the taurine, ?-aminoadipic acid (aaa), glycine, cysteine, NH4, and arginine (Arg) levels were significantly higher compared to control. Significant risk variables for the NIHF group and odds coefficients were obtained using a binary logistic regression method. The respective odds ratios and 95% confidence intervals for the risk variables phosphoserine...

Amniotic fluid amino acid levels in non-immune hydrops fetalis: a case-control study

Erdemo?lu,M.; Kuyumcuoglu,U.; Guzel,A.I.; Celik,Y.; Kale,E.
Fonte: Associação Brasileira de Divulgação Científica Publicador: Associação Brasileira de Divulgação Científica
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/07/2011 Português
Relevância na Pesquisa
66.07%
In a prospective case-control study, we compared the amniotic fluid amino acid levels in non-immune hydrops fetalis (NIHF) and normal fetuses. Eighty fetuses underwent amniocentesis for different reasons at the prenatal diagnosis unit of the Department of Obstetrics and Gynecology, Faculty of Medicine, Dicle University. Forty of these fetuses were diagnosed with NIHF. The study included 40 women each in the NIHF (mean age: 27.69 ± 4.56 years) and control (27.52 ± 5.49 years) groups, who had abnormal double- or triple-screening test values with normal fetuses with gestational ages of 23.26 ± 1.98 and 23.68 ± 1.49 weeks at the time of sample collection, respectively. Amniotic fluid amino acid concentrations (intra-assay variation: 2.26-7.85%; interassay variation: 3.45-8.22%) were measured using EZ:faast kits (EZ:faast GC/FID free (physiological) amino acid kit; Phenomenex, USA) by gas chromatography. The standard for quantitation was a mixture of free amino acids from Phenomenex. The levels of 21 amino acids were measured. The mean phosphoserine and serine levels were significantly lower in the NIHF group, while the taurine, ?-aminoadipic acid (aaa), glycine, cysteine, NH4, and arginine (Arg) levels were significantly higher compared to control. Significant risk variables for the NIHF group and odds coefficients were obtained using a binary logistic regression method. The respective odds ratios and 95% confidence intervals for the risk variables phosphoserine...

Purification of an L-amino acid oxidase from Bungarus caeruleus (Indian krait) venom

More,SS; Kiran,KM; Veena,SM; Gadag,JR
Fonte: Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP, Universidade Estadual Paulista - UNESP Publicador: Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP, Universidade Estadual Paulista - UNESP
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/01/2010 Português
Relevância na Pesquisa
66.02%
Snake venoms are rich in enzymes such as phospholipase A2, proteolytic enzymes, hyaluronidases and phosphodiesterases, which are well characterized. However, L-amino acid oxidase (LAO EC.1.4.3.2) from snake venoms has not been extensively studied. A novel L-amino acid oxidase from Bungarus caeruleus venom was purified to homogeneity using a combination of ion-exchange by DEAE-cellulose chromatography and gel filtration on Sephadex® G-100 column. The purified monomer of LAO showed a molecular mass of 55 ±1 kDa estimated by SDS-PAGE. The specific activity of purified LAO was 6,230 ± 178 U/min/mg, versus 230 ± 3.0 U/min/mg for the whole desiccated venom, suggesting a 27-fold purification with a 25% yield. Optimal pH and temperature for maximum purified enzyme activity were 6.5 and 37ºC, respectively. Platelet aggregation studies show that purified LAO inhibited ADP-induced platelet aggregation dose-dependently at 0.01 to 0.1 µM with 50% inhibitory concentration (IC50) of 0.04 µM, whereas at a 0.08 µM concentration it did not induce appreciable aggregation on normal platelet-rich plasma (PRP). The purified protein catalyzed oxidative deamination of L-amino acids while the most specific substrate was L-leucine. The purified LAO oxidizes only L-forms...

Purification of an L-amino acid oxidase from Bungarus caeruleus (Indian krait) venom

More,SS; Kiran,KM; Veena,SM; Gadag,JR
Fonte: Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP, Universidade Estadual Paulista - UNESP Publicador: Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP, Universidade Estadual Paulista - UNESP
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/01/2010 Português
Relevância na Pesquisa
66.02%
Snake venoms are rich in enzymes such as phospholipase A2, proteolytic enzymes, hyaluronidases and phosphodiesterases, which are well characterized. However, L-amino acid oxidase (LAO EC.1.4.3.2) from snake venoms has not been extensively studied. A novel L-amino acid oxidase from Bungarus caeruleus venom was purified to homogeneity using a combination of ion-exchange by DEAE-cellulose chromatography and gel filtration on Sephadex® G-100 column. The purified monomer of LAO showed a molecular mass of 55 ±1 kDa estimated by SDS-PAGE. The specific activity of purified LAO was 6,230 ± 178 U/min/mg, versus 230 ± 3.0 U/min/mg for the whole desiccated venom, suggesting a 27-fold purification with a 25% yield. Optimal pH and temperature for maximum purified enzyme activity were 6.5 and 37ºC, respectively. Platelet aggregation studies show that purified LAO inhibited ADP-induced platelet aggregation dose-dependently at 0.01 to 0.1 µM with 50% inhibitory concentration (IC50) of 0.04 µM, whereas at a 0.08 µM concentration it did not induce appreciable aggregation on normal platelet-rich plasma (PRP). The purified protein catalyzed oxidative deamination of L-amino acids while the most specific substrate was L-leucine. The purified LAO oxidizes only L-forms...

Amino Acid Metabolic Origin as an Evolutionary Influence on Protein Sequence in Yeast

de Bivort, Benjamin Lovegren; Perlstein, Ethan O.; Kunes, Samuel M.; Schreiber, Stuart L.
Fonte: Springer-Verlag Publicador: Springer-Verlag
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
66.1%
The metabolic cycle of Saccharomyces cerevisiae consists of alternating oxidative (respiration) and reductive (glycolysis) energy-yielding reactions. The intracellular concentrations of amino acid precursors generated by these reactions oscillate accordingly, attaining maximal concentration during the middle of their respective yeast metabolic cycle phases. Typically, the amino acids themselves are most abundant at the end of their precursor’s phase. We show that this metabolic cycling has likely biased the amino acid composition of proteins across the S. cerevisiae genome. In particular, we observed that the metabolic source of amino acids is the single most important source of variation in the amino acid compositions of functionally related proteins and that this signal appears only in (facultative) organisms using both oxidative and reductive metabolism. Periodically expressed proteins are enriched for amino acids generated in the preceding phase of the metabolic cycle. Proteins expressed during the oxidative phase contain more glycolysis-derived amino acids, whereas proteins expressed during the reductive phase contain more respiration-derived amino acids. Rare amino acids (e.g., tryptophan) are greatly overrepresented or underrepresented...