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Mass Spectrometry and X-ray Diffraction Analysis of Two Crystal Types of Dioclea virgata Lectin: An Antinociceptive Protein Candidate to Structure/Function Analysis

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Abstract

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 β 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.

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Acknowledgments

This work was supported by Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico—FUNCAP, Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—CAPES, National Synchrotron Light Laboratory—LNLS, Brazil. BSC and PD are senior investigators of CNPq.

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Authors and Affiliations

  1. Departamento de Biologia Molecular, Universidade Federal da Paraíba, João Pessoa, Brazil

    Plínio Delatorre, Carlos A. A. Gadelha, Tatiane Santi-Gadelha & Daniel L. Farias

  2. Biomol-Lab, Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, Fortaleza, P.O. Box 6043, Zip Code 60455-970, Brazil

    Bruno A. M. Rocha, Rafael C. Simões, Francisco N. Pereira-Júnior, Helton C. Silva, Eduardo Henrique S. Bezerra, Maria Julia B. Bezerra, Emmanuel S. Marinho, Celso S. Nagano & Benildo S. Cavada

  3. Instituto Superior de Ciências Biomédicas, Universidade Estadual do Ceará, Fortaleza, Brazil

    Ana Maria S. Assreuy & Gabriela F. O. Marques-Domingos

  4. Programa de Pós Graduação em Química e Biotecnologia da Universidade Federal de Alagoas, Maceio, Brazil

    Bruno A. M. Rocha

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  1. Plínio Delatorre
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Correspondence to Benildo S. Cavada.

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Delatorre, P., Rocha, B.A.M., Simões, R.C. et al. Mass Spectrometry and X-ray Diffraction Analysis of Two Crystal Types of Dioclea virgata Lectin: An Antinociceptive Protein Candidate to Structure/Function Analysis. Appl Biochem Biotechnol 164, 741–754 (2011). https://doi.org/10.1007/s12010-011-9170-x

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