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You searched for +publisher:"Universidade Federal do Ceará" +contributor:("Ana Maria Sampaio Assereuy"). One record found.

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1. Maria Luciana Lira de Andrade. Dioclea sclerocarpa recombinant lectin: production, biochemical characterization and vasorelaxing activity.

Degree: PhD, 2015, Universidade Federal do Ceará

Lectins are proteins or glycoproteins with at least one non-catalytic domain, which recognizes and binds specific and reversibly to carbohydrates, without modifying its structure. Although these molecules are ubiquitous in nature, the ones isolated from members of the Leguminosae family are the most extensively studied, with a well-characterized biotechnological potential. Among these, the lectins from species of the Diocleinae subtribe are of special interest since they present a wide spectrum of biological activities despite their high structural similarity. During their synthesis in plant cell, these proteins undergo a post-translational processing that result in the formation of three chains (α, β e γ), which constitute the lectins subunits. Moreover, different combination of these chains can randomly raise a mixture of distinct lectin isoforms, causing an broad variability that can impair their biotechnological exploitation. Thus, the present study aimed to produce a recombinant form of the lectin from Dioclea sclerocarpa seeds (DSL), exclusively constituted by α chain. For this, we constructed a synthetic gene from the lectin amino acid sequence, and successfully expressed it in E. coli (BL21). The best condition for the lectin expression was observed using 0.5 mM IPTG induction and growing for 16 h under a 30 °C temperature. The recombinant DSL (rDSL) was then purified by a Sephadex G-50 affinity chromatography, followed by a 0.1 M glucose elution. The rDSL presented an apparent molecular mass similar to the native protein (25 kDa) and the final yield was about 74 mg of protein per liter of culture medium. The rDSL presented specificity to untreated, trypsinized and papainized rabbit blood, and its specific haemagglutinant activity was 13 x 106 H.U./mg. In addition, the rDSL was inhibited in the presence of the sugars D-mannose, α-methyl-mannoside and melibiose, unlike the native protein, which presented haemagglutinant activity even under Melibiose presence. Using in silico molecular modeling techniques, a structural DSL model was obtained and submitted to molecular dynamics, revealing that the protein structure maintains its stability in aqueous environment. From the obtained model, the lectin sugar binding properties was studied using molecular docking. The rDSL presented an astonishing dose-dependent vasorelaxing effect, relaxing rat aortas up to 100% at 100 Âg/mL concentration. This effect was inhibited at ligand sugar presence (Melibiose), as well as by L-NAME. A dose-dependent edematogenic effect was also exerted by rDSL, as evidenced by the paw edema method. However, at the maximum tested dose (300 Âg per paw) a biphasic edema was observed, with the first symptoms after 3 h of administration and another set of symptoms after 24 h. When compared to control values, the edema volume increased 24 times upon rDSL application and this effect was only partially reverted by Melibiose, and completed inhibited by indometacine and L-NAME. Taken together, these results evidence an… Advisors/Committee Members: Benildo Sousa Cavada, Ana Maria Sampaio Assereuy, Francisco Nascimento Pereira JÃnior, Kyria Santiago do Nascimento, CÃntia Renata Costa Rocha.

Subjects/Keywords: BIOLOGIA MOLECULAR; Lectina de Dioclea sclerocarpa; ProteÃna recombinante; CaracterizaÃÃo bioquÃmica; Atividade vasorrelaxante; InflamaÃÃo; Dioclea sclerocarpa lectin; Recombinant protein; Biochemical characterization; Vasorelaxing effect; Inflammation

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APA (6th Edition):

Andrade, M. L. L. d. (2015). Dioclea sclerocarpa recombinant lectin: production, biochemical characterization and vasorelaxing activity. (Doctoral Dissertation). Universidade Federal do Ceará. Retrieved from http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=17128 ;

Chicago Manual of Style (16th Edition):

Andrade, Maria Luciana Lira de. “Dioclea sclerocarpa recombinant lectin: production, biochemical characterization and vasorelaxing activity.” 2015. Doctoral Dissertation, Universidade Federal do Ceará. Accessed April 10, 2021. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=17128 ;.

MLA Handbook (7th Edition):

Andrade, Maria Luciana Lira de. “Dioclea sclerocarpa recombinant lectin: production, biochemical characterization and vasorelaxing activity.” 2015. Web. 10 Apr 2021.

Vancouver:

Andrade MLLd. Dioclea sclerocarpa recombinant lectin: production, biochemical characterization and vasorelaxing activity. [Internet] [Doctoral dissertation]. Universidade Federal do Ceará 2015. [cited 2021 Apr 10]. Available from: http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=17128 ;.

Council of Science Editors:

Andrade MLLd. Dioclea sclerocarpa recombinant lectin: production, biochemical characterization and vasorelaxing activity. [Doctoral Dissertation]. Universidade Federal do Ceará 2015. Available from: http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=17128 ;

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