The P. aeruginosa heme binding protein PhuS is a heme oxygenase titratable regulator of heme uptake.
The Pseudomonas aeruginosa heme utilization (Phu) system encodes several proteins involved in the acquisition of heme as an iron source. Once internalized, heme is degraded by the iron-regulated heme oxygenase, HemO to biliverdin (BV) IXδ and β. In vitro studies have shown holo-PhuS transfers heme to the iron-regulated HemO. This protein-protein interaction is specific for HemO as PhuS does not interact with the α-regioselective heme oxygenase, BphO. Bacterial genetics and isotopic labeling ((13)C-heme) studies confirmed extracellular heme is converted to (13)C-BVIX δ and β through the catalytic action of HemO. In an effort to further understand the role of PhuS, similar studies were performed on the P. aeruginosa PAO1 ΔphuS and ΔphuS/ΔhemO strains. In contrast to wild-type strain, the absence of PhuS results in extracellular heme uptake and degradation via the catalytic action of HemO and BphO. At low heme concentrations, loss of PhuS leads to inefficient extracellular heme uptake supported by the fact the mRNA levels of PhuR, HemO, and BphO remain elevated when compared to the wild-type PAO1. On increasing extracellular heme concentrations, the elevated levels of PhuR, HemO, and BphO allow "leaky uptake" and degradation of heme via HemO and BphO. Similarly, in the ΔphuS/ΔhemO strain, the higher heme concentrations combined with elevated levels of PhuR and BphO leads to nonspecific heme uptake and degradation by BphO. Thus we propose heme flux into the cell is driven by the catalytic action of HemO with PhuS acting as a "control valve" to regulate extracellular heme flux.
The Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, HSF II, 20 Penn Street, Baltimore, Maryland 21201-1140, United States.
Maura J
MJ
O'Neill
Angela
A
Wilks
D001426:Bacterial Proteins / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry*; D002853:Chromatography, Liquid; D003593:Cytoplasm / Q000737:chemistry*; D006418:Heme / Q000493:pharmacokinetics*; D006419:Heme Oxygenase (Decyclizing) / Q000737:chemistry* / Q000493:pharmacokinetics; D000081062:Heme-Binding Proteins; D006420:Hemeproteins / Q000737:chemistry*; D013058:Mass Spectrometry; D008954:Models, Biological; D015394:Molecular Structure; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000201:enzymology
D001426:Bacterial Proteins; D002352:Carrier Proteins; D000081062:Heme-Binding Proteins; D006420:Hemeproteins; D006418:Heme; D006419:Heme Oxygenase (Decyclizing)