Download MendeleyChaperone-assisted cryo-EM structure of P. aeruginosa PhuR reveals molecular basis for heme binding.
Knejski, P.P., Erramilli, S.K., Kossiakoff, A.A.(2024) Structure 32: 411
- PubMed: 38325368
- DOI: https://doi.org/10.1016/j.str.2024.01.007
- Primary Citation of Related Structures:
8THE - PubMed Abstract:
Pathogenic bacteria, such as Pseudomonas aeruginosa, depend on scavenging heme for the acquisition of iron, an essential nutrient. The TonB-dependent transporter (TBDT) PhuR is the major heme uptake protein in P. aeruginosa clinical isolates. However, a comprehensive understanding of heme recognition and TBDT transport mechanisms, especially PhuR, remains limited. In this study, we employed single-particle cryogenic electron microscopy (cryo-EM) and a phage display-generated synthetic antibody (sAB) as a fiducial marker to enable the determination of a high-resolution (2.5 Å) structure of PhuR with a bound heme. Notably, the structure reveals iron coordination by Y529 on a conserved extracellular loop, shedding light on the role of tyrosine in heme binding. Biochemical assays and negative-stain EM demonstrated that the sAB specifically targets the heme-bound state of PhuR. These findings provide insights into PhuR's heme binding and offer a template for developing conformation-specific sABs against outer membrane proteins (OMPs) for structure-function investigations.
Organizational Affiliation:
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA; Laboratory of Medical Biology, Faculty of Biotechnology, University of Wrocław, 50-383 Wrocław, Poland.
