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, Frank A. Ferrone

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

As part of an effort to understand the interactions in HbS polymerization, we have produced and studied a recombinant triple mutant, D6A(α)/D75Y(α)/E121R(β), and a quadruple mutant comprising the preceding mutation plus the natural genetic mutation of sickle hemoglobin, E6V(β). 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.

Original languageEnglish (US)
Pages (from-to)13479-13487
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number16
DOIs
StatePublished - Apr 19 2002
Externally publishedYes

Fingerprint

Sickle Hemoglobin
Dextrans
Copolymerization
Polymers
Solubility
Polymerization
Dimers
Yeast
Mutation
Hemoglobins
Entropy
Oxygen
Yeasts
hemoglobin polymer

ASJC Scopus subject areas

  • Biochemistry

Cite this

Li, X., Briehl, R. W., Bookchin, R. M., Josephs, R., Wei, B., Manning, J. M., & Ferrone, F. A. (2002). Sickle hemoglobin polymer stability probed by triple and quadruple mutant hybrids. Journal of Biological Chemistry, 277(16), 13479-13487. https://doi.org/10.1074/jbc.M108149200

Sickle hemoglobin polymer stability probed by triple and quadruple mutant hybrids. / Li, Xianfeng; Briehl, Robin W.; Bookchin, Robert M.; Josephs, Robert; Wei, Baoyang; Manning, James M.; Ferrone, Frank A.

In: Journal of Biological Chemistry, Vol. 277, No. 16, 19.04.2002, p. 13479-13487.

Research output: Contribution to journalArticle

Li, X, Briehl, RW, Bookchin, RM, Josephs, R, Wei, B, Manning, JM & Ferrone, FA 2002, 'Sickle hemoglobin polymer stability probed by triple and quadruple mutant hybrids', Journal of Biological Chemistry, vol. 277, no. 16, pp. 13479-13487. https://doi.org/10.1074/jbc.M108149200
Li X, Briehl RW, Bookchin RM, Josephs R, Wei B, Manning JM et al. Sickle hemoglobin polymer stability probed by triple and quadruple mutant hybrids. Journal of Biological Chemistry. 2002 Apr 19;277(16):13479-13487. https://doi.org/10.1074/jbc.M108149200
Li, Xianfeng ; Briehl, Robin W. ; Bookchin, Robert M. ; Josephs, Robert ; Wei, Baoyang ; Manning, James M. ; Ferrone, Frank A. / Sickle hemoglobin polymer stability probed by triple and quadruple mutant hybrids. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 16. pp. 13479-13487.
@article{8400976ffdf5488390494aaf812cbe56,
title = "Sickle hemoglobin polymer stability probed by triple and quadruple mutant hybrids",
abstract = "As part of an effort to understand the interactions in HbS polymerization, we have produced and studied a recombinant triple mutant, D6A(α)/D75Y(α)/E121R(β), and a quadruple mutant comprising the preceding mutation plus the natural genetic mutation of sickle hemoglobin, E6V(β). 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.",
author = "Xianfeng Li and Briehl, {Robin W.} and Bookchin, {Robert M.} and Robert Josephs and Baoyang Wei and Manning, {James M.} and Ferrone, {Frank A.}",
year = "2002",
month = "4",
day = "19",
doi = "10.1074/jbc.M108149200",
language = "English (US)",
volume = "277",
pages = "13479--13487",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "16",

}

TY - JOUR

T1 - Sickle hemoglobin polymer stability probed by triple and quadruple mutant hybrids

AU - Li, Xianfeng

AU - Briehl, Robin W.

AU - Bookchin, Robert M.

AU - Josephs, Robert

AU - Wei, Baoyang

AU - Manning, James M.

AU - Ferrone, Frank A.

PY - 2002/4/19

Y1 - 2002/4/19

N2 - As part of an effort to understand the interactions in HbS polymerization, we have produced and studied a recombinant triple mutant, D6A(α)/D75Y(α)/E121R(β), and a quadruple mutant comprising the preceding mutation plus the natural genetic mutation of sickle hemoglobin, E6V(β). 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.

AB - As part of an effort to understand the interactions in HbS polymerization, we have produced and studied a recombinant triple mutant, D6A(α)/D75Y(α)/E121R(β), and a quadruple mutant comprising the preceding mutation plus the natural genetic mutation of sickle hemoglobin, E6V(β). 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.

UR - http://www.scopus.com/inward/record.url?scp=0037134496&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037134496&partnerID=8YFLogxK

U2 - 10.1074/jbc.M108149200

DO - 10.1074/jbc.M108149200

M3 - Article

VL - 277

SP - 13479

EP - 13487

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 16

ER -