Yeast [FeFe]-hydrogenase-like protein Nar1 binds a [2Fe-2S] cluster.

ABSTRACT: Nar1 is an essential eukaryotic protein proposed to function as an iron-sulphur (Fe/S) cluster trafficking factor in the cytosolic iron-sulphur protein assembly (CIA) machinery. However, such a role has remained unclear due to difficulties in purifying adequate amounts of cofactor-bound protein. The [FeFe]-hydrogenase-like protein has two conserved binding sites for [4Fe-4S] clusters but does not show hydrogenase activity in vivo due to the lack of an active site [2Fe]_H cofactor. Here, we report a new preparation procedure for Nar1 that facilitated studies by UV-vis, EPR, and Mössbauer spectroscopies, along with native mass spectrometry. Nar1 recombinantly produced in E. coli contained a [4Fe-4S] cluster, bound presumably at site 1, along with an unexpected [2Fe-2S

SUBMITTER: Braymer JJ 

PROVIDER: S-EPMC12621388 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature