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Anacardium occidentale Bark Lectin: Purification, Immobilization as an Affinity Model and Influence in the Uptake of Technetium-99M by Rat Adipocytes

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Abstract

Lectins, proteins that recognize carbohydrates, have been immobilized on inert supports and used in the screening or purification of glycoproteins. Anacardium occidentale bark infusion has been used as a hypoglycemic agent in Brazil. The toxicity of natural products may be evaluated determining their capability to alter the biodistribution of technetium-99M (99mTc). This work reports the isolation and characterization of a lectin from A. occidentale bark (AnocBL), its evaluation as an affinity support for glycoprotein isolation and lectin effect on the uptake of 99mTc by rat adipocytes. AnocBL was isolated from 80 % ammonium sulphate supernatant by affinity chromatography on fetuin–agarose. SDS–PAGE showed a single protein band of 47 kDa. The monossacharide l-arabinose and the glycoproteins fetuin, asialofetuin, ovomucoid, casein, thyroglobulin, peroxidase, fetal bovine serum and IgG inhibited the activity. The lectin activity was stable until 70 °C and at a pH range of 3.0–7.5. AnocBL–Sepharose column bound fetuin indicating that the lectin matrix may be used to obtain glycoconjugates of biotechnological interest. In vitro assay revealed that glucose and insulin increase 99mTc uptake by rat adipocytes. AnocBL decreases 99mTc uptake, and this effect was not detected in the presence of glucose. Fetuin inhibited AnocBL effect in all insulin concentrations.

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Acknowledgments

The authors express their gratitude to the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for research grants and fellowship (MISM, PMGP and LCBBC). They are also deeply grateful for the technical assistance of Maria Barbosa Reis da Silva.

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Authors and Affiliations

  1. Departamento de Ciências Domésticas, Alimentos Nutrição e Saúde, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, Brazil

    Maria Inês Sucupira Maciel

  2. Instituto de Ciências Biológicas, Universidade de Pernambuco, Recife, Pernambuco, Brazil

    Maria do Socorro de Mendonça Cavalcanti

  3. Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, Pernambuco, 50670-420, Brazil

    Thiago Henrique Napoleão, Patrícia Maria Guedes Paiva & Luana Cassandra Breitenbach Barroso Coelho

  4. Departamento de Biofísica e Radiobiologia, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil

    Maria Teresa Jansem de Almeida Catanho

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  1. Maria Inês Sucupira Maciel
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  2. Maria do Socorro de Mendonça Cavalcanti
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Correspondence to Luana Cassandra Breitenbach Barroso Coelho.

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Maciel, M.I.S., de Mendonça Cavalcanti, M.d.S., Napoleão, T.H. et al. Anacardium occidentale Bark Lectin: Purification, Immobilization as an Affinity Model and Influence in the Uptake of Technetium-99M by Rat Adipocytes. Appl Biochem Biotechnol 168, 580–591 (2012). https://doi.org/10.1007/s12010-012-9798-1

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