Targeting the selectivity filter to drastically alter the activity and substrate spectrum of a promiscuous metal transporter.

ABSTRACT: d-Block metal transporters play a crucial role in maintaining the homeostasis of life-essential trace elements and are attractive targets for protein engineering aimed to selectively enriching or excluding metals in living organisms. However, systematic efforts to engineer these transporters have been hindered by limited understanding of their transport mechanism and substrate specificity. In this study, we applied a focused-screen approach to human ZIP8, a promiscuous d-block divalent metal transporter, by systematically changing three key residues that form the selectivity filter at the entrance of the transport pathway. Screening a library of 48 constructs using an ICP-MS-based transport assay, we identified variants with significantly altered transport activities and/or s

SUBMITTER: Jiang Y 

PROVIDER: S-EPMC12132181 | biostudies-literature | 2025 May

REPOSITORIES: biostudies-literature