6tt9 | pdb_00006tt9
rTBL Recombinant Lectin From Tepary Bean
Structural highlights
Publication Abstract from PubMedHerein, 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 (Ca(2+) and Mn(2+)) attached to each subunit, confirmed in its 3D structure solved by X-ray diffraction at 1.9 A 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 beta1-6 branched N-glycans' characteristics of cancer cells. The presence of alpha1-6 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. Recombinant Lectin from Tepary Bean (Phaseolus acutifolius) with Specific Recognition for Cancer-Associated Glycans: Production, Structural Characterization, and Target Identification.,Martinez-Alarcon D, Varrot A, Fitches E, Gatehouse JA, Cao M, Pyati P, Blanco-Labra A, Garcia-Gasca T Biomolecules. 2020 Apr 23;10(4). pii: biom10040654. doi: 10.3390/biom10040654. PMID:32340396[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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