Apo YfeA extracted from the E. coli periplasm [unknown]

C.D. Radka, S.L. Labiuk, L.J. DeLucas, S.G. Aller
2019 unpublished
In the structural biology of bacterial substrate-binding proteins (SBPs), a growing number of comparisons between substrate-bound and substrate-free forms of metal atom-binding (cluster A-I) SBPs have revealed minimal structural differences between forms. These observations contrast with SBPs that bind substrates such as amino acids or nucleic acids and may undergo >60 rigid-body rotations. Substrate transfer in these SBPs is described by a Venus flytrap model, although this model may not apply
more » ... model may not apply to all SBPs. In this report, structures are presented of substrate-free (apo) and reconstituted substratebound (holo) YfeA, a polyspecific cluster A-I SBP from Yersinia pestis. It is demonstrated that an apo cluster A-I SBP can be purified by fractionation when co-expressed with its cognate transporter, adding an alternative strategy to the mutagenesis or biochemical treatment used to generate other apo cluster A-I SBPs. The apo YfeA structure contains 111 disordered protein atoms in a mobile helix located in the flexible carboxy-terminal lobe. Metal binding triggers a 15-fold reduction in the solvent-accessible surface area of the metal-binding site and reordering of the 111 protein atoms in the mobile helix. The flexible lobe undergoes a 13.6 rigid-body rotation that is driven by a spring-hammer metal-binding mechanism. This asymmetric rigid-body rotation may be unique to metal atom-binding SBPs (i.e. clusters A-I, A-II and D-IV). research papers Acta Cryst. (2019). D75, 831-840 Radka et al. YfeA 839
doi:10.2210/pdb6q1c/pdb fatcat:pxvddx2ia5edfkfszoo6whkkha