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91.
S. Casal B. Macedo M. B. P. P. Oliveira 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2001,763(1-2)
A method is described for evaluation of fat-soluble vitamin in human adipose tissue with the aim to obtain, accurately and within the shortest analysis time, a time-integrated measure of exposure to vitamins from the diet. Fat tissue was deproteinized with ethanol and extracted with n-hexane. Normal-phase HPLC was performed in a Lichrosorb Si60 column with a gradient of n-hexane–2-propanol at 1 ml/min. Detection was accomplished using a diode-array system (for retinol and β-carotene) in series with a fluorescence detector (α-tocopherol). The method was validated and applied to human adipose tissue in a total of 140 subjects. The mean contents found were 0.43, 0.84, 240.3 μg/g for retinol, β-carotene and α-tocopherol, respectively. The method is sensitive enough for detecting the compounds in 1.6 mg of adipose tissue considering the lowest concentration found. 相似文献
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93.
Timmers LF Ducati RG Sánchez-Quitian ZA Basso LA Santos DS de Azevedo WF 《Journal of molecular modeling》2012,18(2):467-479
Cytidine Deaminase (CD) is an evolutionarily conserved enzyme that participates in the pyrimidine salvage pathway recycling cytidine and deoxycytidine
into uridine and deoxyuridine, respectively. Here, our goal is to apply computational techniques in the pursuit of potential
inhibitors of Mycobacterium tuberculosis CD (MtCDA) enzyme activity. Molecular docking simulation was applied to find the possible hit compounds. Molecular dynamics simulations
were also carried out to investigate the physically relevant motions involved in the protein-ligand recognition process, aiming
at providing estimates for free energy of binding. The proposed approach was capable of identifying a potential inhibitor,
which was experimentally confirmed by IC50 evaluation. Our findings open up the possibility to extend this protocol to different databases in order to find new potential
inhibitors for promising targets based on a rational drug design process. 相似文献
94.
Roney Fontes Guimarães Luciana Rodrigues Camillo Aurizangela Oliveira Sousa Luana Pereira Gonçalves Jamilly Costa Cardoso Macedo Giovano Sousa Siqueira Antônia Marlene Magalhães Barbosa Fátima Cerqueira Alvim Marcio Gilberto Cardoso Costa Carlos Priminho Pirovani 《Tree Genetics & Genomes》2018,14(6):90
Here, we report for the first time the genome-wide identification and expression analysis of the molecular chaperone BiP genes in Citrus. Six genes encoding the conserved protein domain family GPR78/BiP/KAR2 were identified in the genome of Citrus sinensis and C. clementina. Two of them, named here as CsBiP1 and CsBiP2, were classified as true BiPs based on their deduced amino acid sequences. Alignment of the deduced amino acid sequences of CsBiP1 and CsBiP2 with BiP homologs from soybean and Arabidopsis showed that they contain all the conserved functional motifs of BiPs. Analysis of the promoter region of CsBiPs revealed the existence of cis-acting regulatory sequences involved in abiotic, heat-shock, and endoplasmic reticulum (ER) stress responses. Publicly available RNA-seq data indicated that CsBiP1 is abundantly expressed in leaf, flower, fruit, and callus, whereas CsBiP2 expression is rarely detected in any tissues under normal conditions. Comparative quantitative real-time PCR (qPCR) analysis of expression of these genes between C. sinensis grafted on the drought-tolerant “Rangpur” lime (C. limonia) and -sensitive “Flying Dragon” trifoliate orange (Poncirus trifoliata) rootstocks showed that CsBiP1 was upregulated by drought stress on the former but downregulated on the latter, whereas the CsBiP2 mRNA levels were downregulated on drought-stressed “Flying Dragon,” but remained constant on “Rangpur.” CsBiP2 upregulation was only observed in C. sinensis seedlings subjected to osmotic and cold treatments. Taken together, these results indicate the existence of two highly conserved BiP genes in Citrus that are differentially regulated in the different tissues and in response to abiotic stresses. 相似文献
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96.
B. J. Goodfellow A. L. Macedo Pedro Rodrigues Isabel Moura Victor Wray José J. G. Moura 《Journal of biological inorganic chemistry》1999,4(4):421-430
The use of standard 2D NMR experiments in combination with 1D NOE experiments allowed the assignment of 51 of the 58 spin systems of oxidised [3Fe-4S] ferredoxin isolated from Desulfovibrio gigas. The NMR solution structure was determined using data from 1D NOE and 2D NOESY spectra, as distance constraints, and information from the X-ray structure for the spin systems not detected by NMR in torsion angle dynamics calculations to produce a family of 15 low target function structures. The quality of the NMR family, as judged by the backbone r.m.s.d. values, was good (0.80?Å), with the majority of φ/ψ angles falling within the allowed region of the Ramachandran plot. A comparison with the X-ray structure indicated that the overall global fold is very similar in solution and in the solid state. The determination of the solution structure of ferredoxin II (FdII) in the oxidised state (FdIIox) opens the way for the determination of the solution structure of the redox intermediate state of FdII (FdIIint), for which no X-ray structure is available. 相似文献
97.
Two novel proteinases were isolated from resting sorghum seeds and purified 100-fold. The activity of the purified enzymes was completely inhibited by pepstatin A and was unaffected by PMSF, leupeptin, EDTA and E-64 (L-trans-epoxysuccinyl leucylamino 4 guanidino butane), which indicates that they belong to the class of aspartic proteinases. SDS-PAGE and native-PAGE revealed a monomeric 29-kDa enzyme and a heterodimeric 61-kDa enzyme with two S-S linked subunits of 49 and 12 kDa. The proteases have maximum activity at 45 °C and pH 3.5, with haemoglobin as substrate. Activity at 60 °C is higher than at 30 °C. 相似文献
98.
A novel enzyme, the first metalloproteinase purified from a monocotyledonous plant, was extracted from the endosperm of sorghum seedlings and purified to homogeneity by ion exchange chromatography and size exclusion chromatography. SDS-PAGE analysis reveals a dimeric 17-kDa protein with two 8-kDa subunits linked by disulfide bond(s). The enzyme is 97% inhibited by 1 mM EDTA and is unaffected by inhibitors of aspartic, cysteine, and serine proteinases. Its pH optimum is 7.0 with hemoglobin as substrate. 相似文献
99.