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An N-terminal amphipathic helix governs activity and conformational dynamics of Nramp metal transporters

Jafari, M.; Zhao, H.; Zhang, Y.; Wang, T.; Merz, K. M.; Hu, J.

2026-05-18 biochemistry
10.64898/2026.05.17.725737 bioRxiv
Show abstract

Nramps are a family of proton-coupled divalent metal ion transporters that play critical roles in maintaining homeostasis of essential metals. In humans, Nramp2 (DMT1) mediates iron uptake to support both cellular and systemic iron homeostasis, whereas Nramp1 exports metals from phagosomes, contributing to antimicrobial defense. A survey of experimentally solved Nramp structures reveals that an N-terminal helix (A) immediately preceding TM1 is folded only in the outward-facing or outward-facing occluded states, raising the question of its role in Nramp transport. Here, we combined mutagenesis, transport assays, and molecular dynamics simulations to investigate A. Our results show that deletion or substitution of conserved residues in A markedly reduces Fe2+ transport. Simulations indicate that A behaves as an amphipathic helix in the outward-facing state and preferentially stabilizes this conformation over the inward-facing state, thus acting as a key player in controlling conformational equilibrium. This study provides new insights into structural dynamics of Nramps and potentially sheds light on other LeuT-fold transporters.

Published in Journal of Biological Chemistry (predicted rank #2) · training set

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