Reciprocal allosteric modulation of carbon monoxide and warfarin binding to ferrous human serum heme-albumin

PLoS One. 2013;8(3):e58842. doi: 10.1371/journal.pone.0058842. Epub 2013 Mar 21.

Abstract

Human serum albumin (HSA), the most abundant protein in human plasma, could be considered as a prototypic monomeric allosteric protein, since the ligand-dependent conformational adaptability of HSA spreads beyond the immediate proximity of the binding site(s). As a matter of fact, HSA is a major transport protein in the bloodstream and the regulation of the functional allosteric interrelationships between the different binding sites represents a fundamental information for the knowledge of its transport function. Here, kinetics and thermodynamics of the allosteric modulation: (i) of carbon monoxide (CO) binding to ferrous human serum heme-albumin (HSA-heme-Fe(II)) by warfarin (WF), and (ii) of WF binding to HSA-heme-Fe(II) by CO are reported. All data were obtained at pH 7.0 and 25°C. Kinetics of CO and WF binding to the FA1 and FA7 sites of HSA-heme-Fe(II), respectively, follows a multi-exponential behavior (with the same relative percentage for the two ligands). This can be accounted for by the existence of multiple conformations and/or heme-protein axial coordination forms of HSA-heme-Fe(II). The HSA-heme-Fe(II) populations have been characterized by resonance Raman spectroscopy, indicating the coexistence of different species characterized by four-, five- and six-coordination of the heme-Fe atom. As a whole, these results suggest that: (i) upon CO binding a conformational change of HSA-heme-Fe(II) takes place (likely reflecting the displacement of an endogenous ligand by CO), and (ii) CO and/or WF binding brings about a ligand-dependent variation of the HSA-heme-Fe(II) population distribution of the various coordinating species. The detailed thermodynamic and kinetic analysis here reported allows a quantitative description of the mutual allosteric effect of CO and WF binding to HSA-heme-Fe(II).

MeSH terms

  • Allosteric Regulation
  • Carbon Monoxide / metabolism*
  • Ferrous Compounds / chemistry
  • Ferrous Compounds / metabolism*
  • Heme / chemistry
  • Heme / metabolism*
  • Humans
  • Kinetics
  • Protein Binding
  • Serum Albumin / chemistry
  • Serum Albumin / metabolism*
  • Thermodynamics
  • Warfarin / chemistry
  • Warfarin / metabolism*

Substances

  • Ferrous Compounds
  • Serum Albumin
  • Heme
  • Warfarin
  • Carbon Monoxide

Grants and funding

These authors have no support or funding to report.