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Insights into the Structural Features, Conformational Stability and Functional Activity of the Olneya tesota PF2 Lectin

[ Vol. 28 , Issue. 4 ]

Author(s):

Edgar Acedo-Espinoza, Irlanda Lagarda-Diaz*, Rosina Cabrera, Ana M. Guzman-Partida, Amir Maldonado-Arce, María M. Ortega-Nieblas, Lerma Chan-Chan and Luz Vázquez-Moreno*   Pages 403 - 413 ( 11 )

Abstract:


Background: The O. tesota lectin PF2 is a tetrameric protein with subunits of 33 kDa that recognizes only complex carbohydrates, resistant to proteolytic enzymes and has insecticidal activity against Phaseolus beans pest.

Objective: To explore PF2 lectin features at different protein structural levels and to evaluate the effect of temperature and pH on its functionality and conformational stability.

Methods: PF2 lectin was purified by affinity chromatography. Its primary structure was resolved by mass spectrometry and analyzed by bioinformatic tools, including its tertiary structure homology modeling. The effect of temperature and pH on its conformational traits and stability was addressed by dynamic light scattering, circular dichroism, and intrinsic fluorescence. The hemagglutinating activity was evaluated using a suspension of peripheral blood erythrocytes.

Results: The proposed PF2 folding comprises a high content of beta sheets. At pH 7 and 25°C, the hydrodynamic diameter (Dh) was found to be 12.3 nm which corresponds to the oligomeric native state of PF2 lectin. Dh increased under the other evaluated pH and temperature conditions, suggesting protein aggregation. At basic pH, PF2 exhibited low conformational stability. The native PF2 (pH 7) retained its full hemagglutinating activity up to 45°C and exhibited one transition state with a melting temperature of 76.8°C.

Conclusion: PF2 showed distinctive characteristics found in legume lectins. The pH influences the functionality and conformational stability of the protein. PF2 lectin displayed a relatively narrow thermostability to the loss of secondary structure and hemagglutinating activity.

Keywords:

Circular dichroism, dynamic light scattering, hemagglutination, homology modeling, intrinsic fluorescence, PF2 lectin, pH, thermostability.

Affiliation:

Departamento de Investigaciones Científicas y Tecnológicas, Universidad de Sonora, Hermosillo, Sonora 83000, CONACyT -Universidad de Sonora, Sonora, Centro de Investigación y Desarrollo en Agrobiotecnología Alimentaria (Consortium between Centro de Investigación en Alimentación y Desarrollo and Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco), Pachuca Ciudad del Conocimiento y la Cultura, San Agustín Tlaxiaca, Hidalgo 42163, CONACyT- Centro de Investigación en Alimentación y Desarrollo, A.C., Hermosillo, Sonora 83304, Departamento de Física, Universidad de Sonora, Hermosillo, Sonora 83000, Departamento de Investigaciones Científicas y Tecnológicas, Universidad de Sonora, Hermosillo, Sonora 83000, CONACyT -Universidad de Sonora, Sonora, CONACyT- Centro de Investigación en Alimentación y Desarrollo, A.C., Hermosillo, Sonora 83304

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