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Raniolo-Limongelli_2023_Elsevier_JMB
Journal article

Uracil/H+ symport by FurE refines aspects of the rocking-bundle mechanism of APC-type transporters

  • Zantza, Iliana Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis, Greece
  • Pyrris, Yiannis Department of Biology, National and Kapodistrian University of Athens, Panepistimiopolis, Greece
  • Raniolo, Stefano ORCID Euler Institute (EUL), Università della Svizzera italiana, Switzerland
  • Papadaki, Georgia F. Department of Biology, National and Kapodistrian University of Athens, Panepistimiopolis, Greece
  • Lambrinidis, George Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis, Greece
  • Limongelli, Vittorio ORCID Euler Institute (EUL), Università della Svizzera italiana, Switzerland ; Department of Pharmacy, University of Naples “Federico II”, Naples, Italy
  • Diallinas, George Department of Biology, National and Kapodistrian University of Athens, Panepistimiopolis, Greece ; Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Heraklion, Greece
  • Mikros, Emmanuel Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis, Greece ; Athena Research and Innovation Center in Information Communication & Knowledge Technologies, Marousi, Greece
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  • 2023
Published in:
  • Journal of molecular biology. - 2023, vol. 435, no. 19, p. 168226
Aspergillus nidulans
NCS1 transporters
FurE nucleobase-proton symporter
Funnel metadynamics
Mechanism of function
English Transporters mediate the uptake of solutes, metabolites and drugs across the cell membrane. The eukaryotic FurE nucleobase/H+ symporter of Aspergillus nidulans has been used as a model protein to address structure–function relationships in the APC transporter superfamily, members of which are characterized by the LeuT-fold and seem to operate by the so-called ‘rocking-bundle’ mechanism. In this study, we reveal the binding mode, translocation and release pathway of uracil/H+ by FurE using path collective variable, funnel metadynamics and rational mutational analysis. Our study reveals a stepwise, induced-fit, mechanism of ordered sequential transport of proton and uracil, which in turn suggests that FurE, functions as a multi-step gated pore, rather than employing ‘rocking’ of compact domains, as often proposed for APC transporters. Finally, our work supports that specific residues of the cytoplasmic N-tail are involved in substrate translocation, in line with their essentiality for FurE function.
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Language
  • English
Classification
Medicine
License
CC BY-NC-ND
Open access status
green
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Persistent URL
https://n2t.net/ark:/12658/srd1329319
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