RESUMEN
Goat whey is normally discarded in the milk processing industry. However, several studies have addressed its biological properties and possible use in human or animal diet. The present study aimed to analysis the protein profile of goat whey to evaluate its possible oxidant, antioxidant, antibacterial, antitumour, and cytotoxic activities in vitro against human erythrocytes. Goat whey was skimmed, and crude protein extract (CPE) was obtained. Next, protein fractions (F) were obtained using ammonium sulphate precipitation method. The proteins were characterized by SDS-PAGE, two-dimensional electrophoresis and soluble protein measurements. No significant differences were observed in protein profile of CPE, F 30-60% and F 60-90%. The highest protein content was found in F 60-90% (0.41mgP/mL). All samples, except F 0-30% showed bacteriostatic activity against different bacterial strains. Only CPE at a concentration of 1000µg/mL was haemolytic against human erythrocytes. Oxidant activity against erythrocytes was not observed. Antioxidant activity was observed only for CPE. Cytotoxicity against C6 rat glioma cell line that was performed with CPE revealed tumour cell death>70% at concentrations of 0.05 and 0.1µg/mL. These results demonstrate at first time that CPE may be used as an antioxidant, bacteriostatic and cytotoxic compound against tumour cells.
Asunto(s)
Antibacterianos/metabolismo , Antineoplásicos/metabolismo , Leche/metabolismo , Proteómica , Proteína de Suero de Leche/metabolismo , Animales , Antibacterianos/farmacología , Antineoplásicos/farmacología , Bacterias/efectos de los fármacos , Línea Celular Tumoral , Eritrocitos/citología , Eritrocitos/efectos de los fármacos , Cabras , Hemólisis/efectos de los fármacos , Humanos , Ratas , Proteína de Suero de Leche/farmacologíaRESUMEN
The anti-tumor effects of a newly-discovered lectin, isolated from okra, Abelmoschus esculentus (AEL), were investigated in human breast cancer (MCF7) and skin fibroblast (CCD-1059 sk) cells. AEL induced significant cell growth inhibition (63 %) in MCF7 cells. The expression of pro-apoptotic caspase-3, caspase-9, and p21 genes was increased in MCF7 cells treated with AEL, compared to those treated with controls. In addition, AEL treatment increased the Bax/Bcl-2 ratio in MCF7 cells. Flow cytometry also indicated that cell death (72 %) predominantly occurred through apoptosis. Thus, AEL in its native form promotes selective antitumor effects in human breast cancer cells and may represent a potential therapeutic to combat human breast cancer.
Asunto(s)
Abelmoschus/química , Antineoplásicos/farmacología , Apoptosis , Células Epiteliales/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Lectinas/farmacología , Antineoplásicos/aislamiento & purificación , Caspasas/análisis , Línea Celular Tumoral , Células Epiteliales/fisiología , Fibroblastos/fisiología , Humanos , Lectinas/aislamiento & purificaciónRESUMEN
The lectin from seeds of Dioclea virgata (DvirL) was purified in a single step affinity chromatography, sequenced by tandem mass spectrometry and submitted to crystallization and biological experiments. DvirL has a molecular mass of 25,412 ± 2 Da and the chains ß and γ has 12,817 Da ± 2 and 12,612 Da ± 2, respectively. Primary sequence determination was assigned by tandem mass spectrometry and revealed a protein with 237 amino acids and 87% of identify with ConA. The protein crystals were obtained native and complexed with X-Man using vapor-diffusion method at a constant temperature of 293 K. A complete X-ray dataset was collected at 1.8 Å resolution. DvirL crystals were found to be orthorhombic, belonging to the space group I222, with a unit cell parameters a = 647.5 Å, b = 86.6 Å, c = 90.2 Å. Molecular replacement search found a solution with a correlation coefficient of 77.1% and an R(factor) of 44.6%. The present study also demonstrated that D. virgata lectin presents edematogenic and antinociceptive activities in rodents electing this protein as a candidate to structure/function analysis.
Asunto(s)
Analgésicos/química , Dioclea/química , Lectinas de Plantas/química , Secuencia de Aminoácidos , Analgésicos/aislamiento & purificación , Analgésicos/farmacología , Animales , Cristalización , Edema/tratamiento farmacológico , Humanos , Masculino , Espectrometría de Masas , Ratones , Datos de Secuencia Molecular , Mapeo Peptídico , Lectinas de Plantas/aislamiento & purificación , Lectinas de Plantas/farmacología , Semillas/química , Alineación de Secuencia , Difracción de Rayos XRESUMEN
The effects of a lectin (AaL) from seeds of Araucaria angustifolia were investigated in the model of rat paw edema. In vivo anti-and pro-inflammatory activities, role of sugar residues, inflammatory mediators and systemic toxicity were assessed. Intravenous injection of AaL (0.1-1 mg/kg) dose-dependently inhibited the dextran-induced increase in edema and vascular permeability, which were prevented by association of the lectin with its binding sugar N-acetyl-glucosamine (Glyc-Nac). AaL also significantly inhibited edema induced by serotonin (18%) and compound 48/80 (33%), but not edema induced by histamine. In contrast, when applied by the s.c. route, AaL evoked a paw edema that peaked 1 h later and was partially prevented by association with Glyc-Nac (59%) or by prior i.v. administration of the lectin itself (38.8%). This AaL edematogenic activity was significantly inhibited by pentoxifylline (44.4%) or dexamethasone (51%) and also by depletion of rat paw mast cells (45.6%), but not by L-N-nitro-arginine methyl ester or indomethacin, excluding involvement of nitric oxide and prostaglandins. Treatment of animals with a single anti-inflammatory dose of AaL (1 mg/kg, i.v.) for 7 days did not affect rat corporal mass, liver, kidney, spleen or stomach wet weight, blood leukocyte count, and urea, creatinine or serum transaminase activity. Systemic toxicity was apparent only at much higher doses (LD50=88.3 mg/kg) than those required for the anti-inflammatory effect. Summarizing, AaL exerts anti-and pro-edematogenic actions via interaction with its specific lectin domain. These actions may share a common pathway involving either activation or inhibition of inflammatory mediators from resident mast cells.
Asunto(s)
Antiinflamatorios/farmacología , Quitina/metabolismo , Mastocitos/fisiología , Lectinas de Plantas/farmacología , Semillas/química , Tracheophyta/química , Enfermedad Aguda , Animales , Permeabilidad Capilar/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Edema/prevención & control , Histamina/farmacología , Masculino , Pentoxifilina/farmacología , Ratas , Ratas Wistar , Serotonina/farmacología , p-Metoxi-N-metilfenetilamina/farmacologíaRESUMEN
The crystal structure of Canavalia maritima lectin (ConM) complexed with trehalose and maltose revealed relevant point mutations in ConA-like lectins. ConM with the disaccharides and other ConA-like lectins complexed with carbohydrates demonstrated significant differences in the position of H-bonds. The main difference in the ConM structure is the replacement of Pro202 by Ser202, a residue that promotes the approximation of Tyr12 to the carbohydrate-binding site. The O-6' of the second glucose ring in maltose interacts with Tyr12, while in trehalose the interaction is established by the O-2' and Tyr12, explaining the higher affinity of ConM for disaccharides compared to monosaccharides.