Iron-mediated oxidation induces conformational changes within the redox-sensing protein HbpS.
HbpS is an extracellular oligomeric protein, which has been shown to act in concert with the two-component system SenS-SenR during the sensing of redox stress. HbpS can bind and degrade heme under oxidative stress conditions, leading to a free iron ion. The liberated iron is subsequently coordinated on the protein surface. Furthermore, HbpS has been shown to modulate the phosphorylation state of the sensor kinase SenS as, in the absence of oxidative stress conditions, HbpS inhibits SenS autophosphorylation whereas the presence of heme or iron ions and redox-stressing agents enhances it. Using HbpS wild type and mutants as well as different biochemical and biophysical approaches, we show that iron-mediated oxidative stress induces both secondary structure and overall intrinsic conformational changes within HbpS. We demonstrate in addition that HbpS is oxidatively modified, leading to the generation of highly reactive carbonyl groups and tyrosine-tyrosine bonds. Further examination of the crystal structure and subsequent mutational analyses allowed the identification of the tyrosine residue participating in dityrosine formation, which occurs between two monomers within the octomeric assembly. Therefore, it is proposed that oxidative modifications causing structural and conformational changes are responsible for the control of SenS and hence of the HbpS-SenS-SenR signaling cascade.
Ortiz de Orué Lucana
Darío
D
Department of Applied Genetics of Microorganisms, Faculty of Biology/Chemistry, University of Osnabrück, 49069 Osnabrück, Germany. ortiz@biologie.uni-osnabrueck.de
Roscher
Mareike
M
Honigmann
Alf
A
Schwarz
Julia
J
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D007501:Iron / Q000494:pharmacology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D011487:Protein Conformation / Q000187:drug effects; D013302:Streptomyces / Q000235:genetics; D014158:Transcription, Genetic; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D001426:Bacterial Proteins; D014443:Tyrosine; C007543:dityrosine; D007501:Iron