info:eu-repo/semantics/article
Ancestral-sequence reconstruction unveils the structural basis of function in mammalian FMOs
Fecha
2020-01Registro en:
Nicoll, Callum R.; Bailleul, Gautier; Fiorentini, Filippo; Mascotti, María Laura; Fraaije, Marco Wilhelmus; et al.; Ancestral-sequence reconstruction unveils the structural basis of function in mammalian FMOs; Nature; Nature Structural and Molecular Biology; 27; 1; 1-2020; 14-24
1545-9993
1545-9985
CONICET Digital
CONICET
Autor
Nicoll, Callum R.
Bailleul, Gautier
Fiorentini, Filippo
Mascotti, María Laura
Fraaije, Marco Wilhelmus
Mattevi, Andrea
Resumen
Flavin-containing monooxygenases (FMOs) are ubiquitous in all domains of life and metabolize a myriad of xenobiotics, including toxins, pesticides and drugs. However, despite their pharmacological importance, structural information remains bereft. To further our understanding behind their biochemistry and diversity, we used ancestral-sequence reconstruction, kinetic and crystallographic techniques to scrutinize three ancient mammalian FMOs: AncFMO2, AncFMO3-6 and AncFMO5. Remarkably, all AncFMOs could be crystallized and were structurally resolved between 2.7- and 3.2-Å resolution. These crystal structures depict the unprecedented topology of mammalian FMOs. Each employs extensive membrane-binding features and intricate substrate-profiling tunnel networks through a conspicuous membrane-adhering insertion. Furthermore, a glutamate–histidine switch is speculated to induce the distinctive Baeyer–Villiger oxidation activity of FMO5. The AncFMOs exhibited catalysis akin to human FMOs and, with sequence identities between 82% and 92%, represent excellent models. Our study demonstrates the power of ancestral-sequence reconstruction as a strategy for the crystallization of proteins.