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Related Experiment Videos

Sickle hemoglobin polymer melting in high concentration phosphate buffer.

J G Louderback1, S K Ballas, D B Kim-Shapiro

  • 1Department of Physics, Wake Forest University, Winston-Salem, North Carolina 27109-7507, USA.

Biophysical Journal
|March 30, 1999
PubMed
Summary

Carbon monoxide (CO) binding to sickle hemoglobin (HbS) and HbS polymer melting were studied. CO binds to HbS polymers before they depolymerize, revealing insights into sickle cell disease mechanisms.

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Area of Science:

  • Biophysics
  • Hematology
  • Polymer Science

Background:

  • Sickle cell hemoglobin (HbS) forms polymers under deoxygenated conditions, leading to red blood cell sickling.
  • Understanding the kinetics of HbS polymerization and depolymerization is crucial for developing effective treatments for sickle cell disease.

Purpose of the Study:

  • To investigate the mechanism and kinetics of sickle hemoglobin polymer melting.
  • To determine the rate of carbon monoxide (CO) binding to deoxygenated HbS (deoxy-HbS) in the polymer phase and in solution.

Main Methods:

  • Transmission spectroscopy (optical absorption and turbidity) was used to monitor CO binding and HbS polymer melting simultaneously.
  • Argon-saturated and CO-saturated phosphate buffers were employed for rapid mixing experiments.

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Main Results:

  • Conversion of deoxy-HbS in the polymer phase to the HbS-CO adduct occurred at an average rate of 5.8 s-1, proceeding through an intermediate species.
  • The intermediate species grew in with a rate of 36 s-1, suggesting CO binding to the polymer phase prior to melting.
  • Normal adult hemoglobin (HbA) conversion to HbA-CO adduct showed a faster rate of 83 s-1 under identical conditions.

Conclusions:

  • CO binding to HbS polymers precedes and likely facilitates polymer melting (depolymerization).
  • The findings provide critical kinetic data on HbS polymer dynamics, offering potential therapeutic targets.
  • This study elucidates a key step in the transition of HbS from a polymerized to a depolymerized state.