Ancestral-sequence reconstruction unveils the structural basis of function in mammalian FMOs

Callum R Nicoll, Gautier Bailleul, Filippo Fiorentini, María Laura Mascotti, Marco W Fraaije*, Andrea Mattevi

*Bijbehorende auteur voor dit werk

Onderzoeksoutput: ArticleAcademicpeer review

30 Citaten (Scopus)
254 Downloads (Pure)


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.

Originele taal-2English
Pagina's (van-tot)14-24
Aantal pagina's11
TijdschriftNature Structural & Molecular Biology
Nummer van het tijdschrift1
Vroegere onlinedatum23-dec.-2019
StatusPublished - 2019

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