Conserved hydrophilic checkpoints tune FocA-mediated formate:H<sup>+</sup> symport.

Tüting, Christian; Janson, Kevin; Kammel, Michelle; Ihling, Christian; Lorenz, Jana; Kyrilis, Fotis L; Hamdi, Farzad; Erdmann, Christopher et al. · Nat Commun · 2025

basic_science · Level V

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Abstract

FocA belongs to the widespread, evolutionarily ancient formate-nitrite transporter (FNT) family of pentameric anion channels and translocates formic acid bidirectionally. Here, we identify compartmentalized polarity distribution across the complete FocA pore structure - resolved at 2.56 Å - mirrored against a two-fold axis with H209 at its center. A FocA-H209N variant that exhibits an efflux-only channel-like function in vivo reveals a density consistent with formate located directly at N209, abolishing the channel's amphiphilicity. Pyruvate formate-lyase, which generates formate, orients at the cytoplasmic face where formate delivery is regulated by conformational changes in the FocA vestibule. Comparisons with other FNTs suggest a tuning mechanism of formate-specific transport via checkpoints enriched in hydrophilic residues.

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