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Sickle hemoglobin polymer stability probed by triple and quadruple mutant hybrids
Journal article   Open access   Peer reviewed

Sickle hemoglobin polymer stability probed by triple and quadruple mutant hybrids

Xianfeng Li, Robin W Briehl, Robert M Bookchin, Robert Josephs, Baoyang Wei, James M Manning and Frank A Ferrone
The Journal of biological chemistry, v 277(16), pp 13479-13487
19 Apr 2002
DOI: https://doi.org/10.1074/jbc.M108149200
PMID: 11782463
Featured in Collection :   UN Sustainable Development Goals @ Drexel
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https://doi.org/10.1074/jbc.M108149200View
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Abstract

Hemoglobin, Sickle - chemistry Electrophoresis, Polyacrylamide Gel Hemoglobin A - genetics Humans Isoelectric Focusing Hemoglobin, Sickle - genetics Chromatography, High Pressure Liquid Models, Statistical Recombinant Fusion Proteins - chemistry Plasmids - metabolism Oxygen - metabolism Dose-Response Relationship, Drug Hemoglobin A - chemistry Spectrometry, Mass, Electrospray Ionization Time Factors Kinetics Mutation Circular Dichroism Dimerization
As part of an effort to understand the interactions in HbS polymerization, we have produced and studied a recombinant triple mutant, D6A(alpha)/D75Y(alpha)/E121R(beta), and a quadruple mutant comprising the preceding mutation plus the natural genetic mutation of sickle hemoglobin, E6V(beta). These recombinant hemoglobins expressed in yeast were extensively characterized, and their structure and oxygen binding cooperativity were found to be normal. Their tetramer-dimer dissociation constants were within a factor of 2 of HbA and HbS. Polymerization of these mutants mixed with HbS was investigated by a micromethod based on volume exclusion by dextran. The elevated solubility of mixtures of HbS with HbA and HbF in dextran could be accurately predicted without any variable parameters. Relative to HbS, the copolymerization probability of the quadruple mutant/HbS hybrid was found to be 6.2, and the copolymerization probability for the triple mutant/HbS hybrid was 0.52. The pure quadruple mutant had a solubility slightly above that of its hybrid with HbS. One way to explain these results is to require significant cis-trans differences in the polymer and that HbA assemble above 42.5 g/dl. A second way to explain these data is by the modification of motional freedom, thereby changing vibrational entropy in the polymer.

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