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TitleConserved hydrophilic checkpoints tune FocA-mediated formate:H symport.
Journal, issue, pagesNat Commun, Vol. 16, Issue 1, Page 9476, Year 2025
Publish dateOct 27, 2025
AuthorsChristian Tüting / Kevin Janson / Michelle Kammel / Christian Ihling / Jana Lorenz / Fotis L Kyrilis / Farzad Hamdi / Christopher Erdmann / Andrea Sinz / R Gary Sawers / Panagiotis L Kastritis /
PubMed AbstractFocA belongs to the widespread, evolutionarily ancient formate-nitrite transporter (FNT) family of pentameric anion channels and translocates formic acid bidirectionally. Here, we identify ...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.
External linksNat Commun / PubMed:41145500 / PubMed Central
MethodsEM (single particle)
Resolution2.56 - 2.97 Å
Structure data

51034

9g49

EMDB-51034, PDB-9g49:
Cryo-EM reconstruction of the full-length E. coli transmembrane formate transporter FocA
Method: EM (single particle) / Resolution: 2.56 Å

51035

9g4d

EMDB-51035, PDB-9g4d:
Cryo-EM reconstruction of the full-length H209N mutant E. coli transmembrane formate transporter FocA
Method: EM (single particle) / Resolution: 2.97 Å

52595

9i3k

EMDB-52595, PDB-9i3k:
Asymmetric cryo-EM reconstruction of the full-length E. coli transmembrane formate transporter FocA
Method: EM (single particle) / Resolution: 2.87 Å

Chemicals

ChemComp-HOH:
WATER

Source
  • escherichia coli k-12 (bacteria)
KeywordsMEMBRANE PROTEIN / FNT-transporter / FocA / formate / H209N

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