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Giới thiệu nội dung

Recombinant hemoglobin ẞG83C-F41Y

Authors:

Corinne Vasseur-Godbillon, Sarata C. Sahu, Elisa Domingues, Christophe Fablet, Janel L. Giovannelli, Tsuey Chyi Tam, Nancy T. Ho, Chien Ho, Michael C. Marden, and Véronique Baudin-Creuza

Field:

Biochemistry, Molecular Biology

Document Content:

This document describes the engineering and characterization of a recombinant human hemoglobin (Hb) mutant, rHb ẞG83C-F41Y, designed as a potential blood substitute. This octameric protein is formed by disulfide bridges between tetramers, increasing its molecular size to overcome the vasoactivity issues associated with smaller acellular hemoglobin solutions. The study details the protein’s stability, oxygen binding properties, CO rebinding kinetics, and its behavior in various physiological conditions. The research highlights the stable octameric structure, its resistance to dissociation and degradation, and its potential for therapeutic applications.

Detailed Table of Contents:

(The provided text does not contain a structured table of contents. The following is a breakdown of the main sections based on the document’s content.)

  • Introduction and Background
  • Keywords
  • Correspondence
  • Abstract
  • Abbreviations
  • Results
  • Stability of the ẞG83C-F41Y oligomers
    • Concentration dependent dissociation equilibrium
    • Auto-oxidation
  • Secondary structure and thermal stability
  • Reaction with Hp
  • Stability in fresh plasma
  • Study of disulfide bridge kinetics
  • Octamer to tetramer transition
  • Disulfide bridge formation between rHb BG83C-F41Y tetramers
  • Hybridization with HbA
  • Functional studies
    • CO rebinding kinetics
    • Oxygen-binding properties of octameric rHbs
  • 1H-NMR Studies
  • Cysteine reactivity
  • Acknowledgements
  • Experimental procedures
    • Hemoglobin expression and purification
    • Auto-oxidation kinetics
    • Thermal denaturation
    • Interaction of octamers with haptoglobin
    • Stability of octamers in fresh plasma
    • Disulfide reduction kinetics of the oligomeric rHb BG83C-F41Y
    • Disulfide bridge formation of rHb BG83C-F41Y
    • CO recombination kinetics
    • Oxygen-binding measurements
  • References