Redox-dependent axial ligand replacement and its functional significance in heme-bound iron regulatory proteins

J Inorg Biochem. 2018 May:182:238-248. doi: 10.1016/j.jinorgbio.2018.01.007. Epub 2018 Jan 9.

Abstract

Iron regulatory proteins (IRPs), regulators of iron metabolism in mammalian cells, control the translation of proteins involved in iron uptake, storage and utilization by binding to specific iron-responsive element (IRE) sequences of mRNAs. Two homologs of IRPs (IRP1 and IRP2) have a typical heme regulatory motif (HRM), a consensus sequence found in "heme-regulated proteins". However, specific heme binding to HRM has been reported only for IRP2, which is essential for oxidative modification and loss of binding to target mRNAs. In this paper, we confirmed that IRP1 also specifically binds two molar equivalents of heme, and found that the absorption and resonance Raman spectra of heme-bound IRP1 were quite similar to those of heme-bound IRP2. This shows that the heme environmental structures in IRP1 are close to those of proteins using heme as a regulatory molecule. Pulse radiolysis experiments, however, clearly revealed an axial ligand exchange from Cys to His immediately after the reduction of the heme iron to form a 5-coordinate His-ligated heme in heme-bound IRP2, whereas the 5-coordinate His-ligated heme was not observed after the reduction of heme-bound IRP1. Considering that the oxidative modification is only observed in heme-bound IRP2, but not IRP1, probably owing to the structural flexibility of IRP2, we propose that the transient 5-coordinate His-ligated heme is a prerequisite for oxidative modification of heme-bound IRP2, which functionally differentiates heme binding of IRP2 from that of IRP1.

Keywords: IRP; Ligand exchange; Oxidation; Pulse radiolysis; Resonance Raman.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Heme / chemistry
  • Heme / metabolism
  • Iron / metabolism
  • Iron-Regulatory Proteins / chemistry*
  • Iron-Regulatory Proteins / metabolism*
  • Protein Binding

Substances

  • Iron-Regulatory Proteins
  • Heme
  • Iron