Molecular oxygen migration through the xenon docking sites of human hemoglobin in the R-state

Biochim Biophys Acta. 2016 Sep;1864(9):1110-1121. doi: 10.1016/j.bbapap.2016.06.004. Epub 2016 Jun 7.

Abstract

A nanosecond laser flash-photolysis technique was used to study bimolecular and geminate molecular oxygen (O2) rebinding to tetrameric human hemoglobin and its isolated α and β chains in buffer solutions equilibrated with 1atm of air and up to 25atm of xenon. Xenon binding to the isolated α chains and to the α subunits within tetrameric hemoglobin was found to cause a decrease in the efficiency of O2 escape by a factor of ~1.30 and 3.3, respectively. A kinetic model for O2 dissociation, rebinding, and migration through two alternative pathways in the hemoglobin subunits was introduced and discussed. It was shown that, in the isolated α chains and α subunits within tetrameric hemoglobin, nearly one- and two-third escaping molecules of O2 leave the protein via xenon docking sites, respectively. The present experimental data support the idea that O2 molecule escapes from the β subunits mainly through the His(E7) gate, and show unambiguously that, in the α subunits, in addition to the direct E7 channel, there is at least one alternative escape route leading to the exterior via the xenon docking sites.

Keywords: Hemoglobin; Mercury; Molecular oxygen; Rebinding; Xenon.

MeSH terms

  • Dithiothreitol / chemistry
  • Hemoglobins / chemistry*
  • Hemoglobins / isolation & purification
  • Humans
  • Kinetics
  • Mercuribenzoates / chemistry
  • Molecular Docking Simulation
  • Oxygen / chemistry*
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Multimerization
  • Protein Structure, Secondary
  • Protein Subunits / chemistry*
  • Protein Subunits / isolation & purification
  • Thermodynamics
  • Xenon / chemistry*

Substances

  • 4-mercuribenzoate
  • Hemoglobins
  • Mercuribenzoates
  • Protein Subunits
  • Xenon
  • Oxygen
  • Dithiothreitol