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Recombinant Lectin from Tepary Bean (Phaseolus acutifolius) with Specific Recognition for Cancer-Associated Glycans: Production, Structural Characterization, and Target Identification

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Centro de Investigación y de Estudios Avanzados Unidad Irapuato, Departamento de Biotecnología y Bioquímica, Irapuato 36821, Guanaj uato, Mexico
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University of Grenoble Alpes, CNRS, CERMAV, 38000 Grenoble, France
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Department of Biosciences, Durham University, Durham DH1 3LE, UK
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Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Santiago de Querétaro 76230, Querétaro, Mexico
*
Authors to whom correspondence should be addressed.
Biomolecules 2020, 10(4), 654; https://doi.org/10.3390/biom10040654
Received: 22 March 2020 / Revised: 16 April 2020 / Accepted: 20 April 2020 / Published: 23 April 2020
Herein, we report the production of a recombinant Tepary bean lectin (rTBL-1), its three-dimensional (3D) structure, and its differential recognition for cancer-type glycoconjugates. rTBL-1 was expressed in Pichia pastoris, yielding 316 mg per liter of culture, and was purified by nickel affinity chromatography. Characterization of the protein showed that rTBL-1 is a stable 120 kDa homo-tetramer folded as a canonical leguminous lectin with two divalent cations (Ca2+ and Mn2+) attached to each subunit, confirmed in its 3D structure solved by X-ray diffraction at 1.9 Å resolution. Monomers also presented a ~2.5 kDa N-linked glycan located on the opposite face of the binding pocket. It does not participate in carbohydrate recognition but contributes to the stabilization of the interfaces between protomers. Screening for potential rTBL-1 targets by glycan array identified 14 positive binders, all of which correspond to β1-6 branched N-glycans’ characteristics of cancer cells. The presence of α1-6 core fucose, also tumor-associated, improved carbohydrate recognition. rTBL-1 affinity for a broad spectrum of mono- and disaccharides was evaluated by isothermal titration calorimetry (ITC); however, no interaction was detected, corroborating that carbohydrate recognition is highly specific and requires larger ligands for binding. This would explain the differential recognition between healthy and cancer cells by Tepary bean lectins. View Full-Text
Keywords: recombinant lectins; Tepary bean; Pichia pastoris; MGAT5; glycan array; structure; cancer recombinant lectins; Tepary bean; Pichia pastoris; MGAT5; glycan array; structure; cancer
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MDPI and ACS Style

Martínez-Alarcón, D.; Varrot, A.; Fitches, E.; Gatehouse, J.A.; Cao, M.; Pyati, P.; Blanco-Labra, A.; Garcia-Gasca, T. Recombinant Lectin from Tepary Bean (Phaseolus acutifolius) with Specific Recognition for Cancer-Associated Glycans: Production, Structural Characterization, and Target Identification. Biomolecules 2020, 10, 654. https://doi.org/10.3390/biom10040654

AMA Style

Martínez-Alarcón D, Varrot A, Fitches E, Gatehouse JA, Cao M, Pyati P, Blanco-Labra A, Garcia-Gasca T. Recombinant Lectin from Tepary Bean (Phaseolus acutifolius) with Specific Recognition for Cancer-Associated Glycans: Production, Structural Characterization, and Target Identification. Biomolecules. 2020; 10(4):654. https://doi.org/10.3390/biom10040654

Chicago/Turabian Style

Martínez-Alarcón, Dania, Annabelle Varrot, Elaine Fitches, John A. Gatehouse, Min Cao, Prashant Pyati, Alejandro Blanco-Labra, and Teresa Garcia-Gasca. 2020. "Recombinant Lectin from Tepary Bean (Phaseolus acutifolius) with Specific Recognition for Cancer-Associated Glycans: Production, Structural Characterization, and Target Identification" Biomolecules 10, no. 4: 654. https://doi.org/10.3390/biom10040654

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