title: Folding and homodimerization of wheat germ agglutinin.
abstract: Wheat germ agglutinin (WGA) is emblematic of proteins that specialize in the recognition of carbohydrates. It was the first lectin reported to have a capacity for discriminating between normal and malignant cells. Since then, it has become a preferred model for basic research and is frequently considered in the development of biomedical and biotechnological applications. However, the molecular basis for the structural stability of this homodimeric lectin remains largely unknown, a situation that limits the rational manipulation and modification of its function. In this work we performed a thermodynamic characterization of WGA folding and self-association processes as a function of pH and temperature by using differential scanning and isothermal dilution calorimetry. WGA is monomeric at pH 2, and one of its four hevein-like domains is unfolded at room temperature. Under such conditions, the agglutinin exhibits a fully reversible thermal unfolding that consists of three two-state transitions. At higher pH values, the protein forms weak, nonobligate dimers. This behavior contrasts with that observed for the other plant lectins studied thus far, which form strong, obligate oligomers, indicating a distinctly different molecular basis for WGA function. For dimer formation, the four domains must be properly folded. Nevertheless, depending on the solution conditions, self-association may be coupled with folding of the labile domain. Therefore, dimerization may proceed as a rigid-body-like association or a folding-by-binding event. This hybrid behavior is not seen in other plant lectins. The emerging molecular picture for the WGA assembly highlights the need for a reexamination of existing ligand-binding data in the literature.
authors.affiliation: Instituto de Química, Universidad Nacional Autónoma de México, Circuito Exterior, Ciudad Universitaria, México, DF, México.
authors.forename: María Del Carmen
authors.initials: Mdel C
authors.lastname: Portillo-Téllez
authors.forename: Martiniano
authors.initials: M
authors.lastname: Bello
authors.forename: Guillermo
authors.initials: G
authors.lastname: Salcedo
authors.forename: Gabriel
authors.initials: G
authors.lastname: Gutiérrez
authors.forename: Virginia
authors.initials: V
authors.lastname: Gómez-Vidales
authors.forename: Enrique
authors.initials: E
authors.lastname: García-Hernández
mesh_terms: D050260:Carbohydrate Metabolism; D008024:Ligands; D017510:Protein Folding; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D058767:Protein Unfolding; D014909:Wheat Germ Agglutinins / Q000737:chemistry*
chemical_list: D008024:Ligands; D014909:Wheat Germ Agglutinins