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Universal Metastability of Sickle Hemoglobin Polymerization
Journal article   Open access   Peer reviewed

Universal Metastability of Sickle Hemoglobin Polymerization

Weijun Weng, Alexey Aprelev, Robin W Briehl and Frank A Ferrone
Journal of molecular biology, v 377(4), pp 1228-1235
04 Apr 2008
PMID: 18308336
url
https://doi.org/10.1016/j.jmb.2008.01.083View
Published, Version of Record (VoR) Open

Abstract

metastability polymers sickle-cell disease crowding
Sickle hemoglobin (HbS) polymerization occurs when the concentration of deoxyHbS exceeds a well-defined solubility. In experiments using sickle hemoglobin droplets suspended in oil, it has been shown that when polymerization ceases the monomer concentration is above equilibrium solubility. We find that the final concentration in uniform bulk solutions (i.e., with negligible boundaries) agrees with the droplet measurements, and both exceed the expected solubility. To measure hemoglobin in uniform solutions, we used modulated excitation of trace amounts of CO in gels of HbS. In this method, a small amount of CO is introduced to a spatially uniform deoxyHb sample, so that less than 2% of the sample is liganded. The liganded fraction is photolyzed repeatedly and the rate of recombination allows the concentration of deoxyHbS in the solution phase to be determined, even if polymers have formed. Both uniform and droplet samples exhibit the same quantitative behavior, exceeding solubility by an amount that depends on the initial concentration of the sample, as well as conditions under which the gel was formed. We hypothesize that the early termination of polymerization is due to the obstruction in polymer growth, which is consistent with the observation that pressing on slides lowers the final monomer concentration, making it closer to solubility. The thermodynamic solubility in free solution is thus achieved only in conditions with low polymer density or under external forces (such as found in sedimentation) that disrupt polymers. Since we find that only about 67% of the expected polymer mass forms, this result will impact any analysis predicated on predicting the polymer fraction in a given experiment.

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Collaboration types
Domestic collaboration
Web of Science research areas
Biochemistry & Molecular Biology
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