Expression, Purification, and Functional Characterization of Tectonin 2 from Laccaria bicolor: A Six-Bladed Beta-Propeller Lectin Specific for O-Methylated Glycans

Therese Wohlschlager (1, 2) , Alexander Titz (3) , Markus Künzler (4, 5) , Annabelle Varrot (6)

1 Institute of Microbiology [Zurich]
2 Universität Salzburg
3 HIPS - Chemical Biology of Carbohydrates, Helmholtz Institute for Pharmaceutical Research Saarland
4 Institute of Medical Microbiology [Zurich]
5 ETH Zürich - Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich]
6 CERMAV - Centre de Recherches sur les Macromolécules Végétales

Tectonins are conserved defense proteins of innate immune systems featuring a β-propeller fold. Tectonin 2 from Laccaria bicolor, Lb-Tec2, is the first fungal representative of the tectonin superfamily that has been described. In-depth characterization revealed a specificity for O-methylated glycans and identified a unique sequence motif and binding site architecture underlying this unusual specificity. This chapter provides information on how to produce and purify recombinant Lb-Tec2, characterize its interaction with O-methylated glycans and demonstrate its biological function.

Mots clés: β-Propeller; Non-self recognition; O-Methylated glycans; Defense effector; Nematotoxic lectin; Innate immunity

Domaines: Biochimie, Biologie Moléculaire

Therese Wohlschlager, Alexander Titz, Markus Künzler, Annabelle Varrot. Expression, Purification, and Functional Characterization of Tectonin 2 from Laccaria bicolor: A Six-Bladed Beta-Propeller Lectin Specific for O-Methylated Glycans. Lectin Purification and Analysis: Methods and Protocols, Methods in Molecular Biology, pp.669-682, 2020, ⟨10.1007/978-1-0716-0430-4_58⟩. ⟨hal-02928062⟩