Separation of asymmetrical hybrid hemoglobins by anaerobic cation-exchange high-performance liquid chromatography

Ip, C Y; Asakura, Toshio

doi:10.1016/0003-2697(84)90029-0

Cited by 8 publications

(3 citation statements)

References 14 publications

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“…In effect separation leads to loss of asymmetric species through conversion to symmetric forms. Attempts to prevent loss of asymmetric tetramers by chemical cross-linking (9 -11), retarding dissociation rate by analysis of Hb under anaerobic conditions in the more stable deoxy form (1,2,(12)(13)(14)(15), or performing separation at cryogenic temperatures (16) have been used successfully; however, these approaches are laborious or resolve the hybrid poorly, and are technically difficult.…”

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confidence: 99%

Mass Spectral Analysis of Asymmetric Hemoglobin Hybrids: Demonstration of Hb FS (α2γβS) in Sickle Cell Disease

Ofori-Acquah

Green²,

Davies

et al. 2001

Analytical Biochemistry

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confidence: 99%

Mass Spectral Analysis of Asymmetric Hemoglobin Hybrids: Demonstration of Hb FS (α2γβS) in Sickle Cell Disease

Ofori-Acquah

Green²,

Davies

et al. 2001

Analytical Biochemistry

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“…out by several research groups (Park, 1970(Park, , 1973Bunn & McDonough, 1974;Bunn, 1981; Ip & Asakura, 1984Anbari et al, 1985). Accurate quantitation in these systems was difficult, if not impossible, due to the fact that the hybrid system was continuously perturbed away from equilibrium during the separation procedures used.…”

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confidence: 99%

“…Accurate quantitation in these systems was difficult, if not impossible, due to the fact that the hybrid system was continuously perturbed away from equilibrium during the separation procedures used. When possible at all, quantitation involved extrapolations to zero separation time (Ip & Asakura, 1984Anbari et al, 1985). Perrella and colleagues circumvented this continuous rearrangement by developing new, low-temperature isoelectric focusing and gel electrophoresis technology (Perrella et al, 1978(Perrella et al, , 1983).…”

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confidence: 99%

Direct and indirect pathways of functional coupling in human hemoglobin are revealed by quantitative low-temperature isoelectric focusing of mutant hybrids

LiCata¹,

Speros²,

Rovida³

et al. 1990

Biochemistry

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Functional energetic coupling within human hemoglobin has been explored by using quantitative analysis of asymmetric mutant hybrid equilibria. Previous work showed that the free energy of cooperativity is largely attributable to alterations in free energy that accompany changing interactions at the interface between alpha 1 beta 1 and alpha 2 beta 2 dimers [Pettigrew et al. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 1849]. However, the issue of how cooperativity-linked sites in the molecule are energetically coupled in manifesting cooperative ligation is still not well delineated. In this paper we address the questions of what types of functional coupling pathways are operational in hemoglobin, what some of their characteristics are, and how they are related to one another. By constructing asymmetric mutant hybrid hemoglobins, we can assay how two structurally identical, symmetrically equivalent sites are energetically coupled in manifesting subunit assembly and/or cooperative ligation. Asymmetric hybrid hemoglobins, i.e., those containing a single modified site, cannot be isolated and must be studied in equilibrium with their symmetric parent molecules. In order to study these asymmetric hybrid equilibria, we have developed new theory and quantitation techniques to augment the low-temperature quenching and isoelectric focusing procedures of Perrella et al. [(1978) Anal. Biochem. 88, 212]. Studies of these mutant hybrid hemoglobins have provided evidence for three distinct types of energetic coupling within the hemoglobin tetramer. All alpha 1 beta 2 interface sites examined are involved in cooperativity-linked indirect coupling. Within the context of this indirect "pathway" there exist two different types of direct long-range coupling. One of these classes of direct long-range pathways is linked to cooperative ligand binding while the other class is not.

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Metal Ion Reconstituted Hybrid Hemoglobins

Venkatesh

Manoharan

Rifkind

1997

Progress in Inorganic Chemistry

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Separation of asymmetrical hybrid hemoglobins by anaerobic cation-exchange high-performance liquid chromatography

Cited by 8 publications

References 14 publications

Mass Spectral Analysis of Asymmetric Hemoglobin Hybrids: Demonstration of Hb FS (α2γβS) in Sickle Cell Disease

Mass Spectral Analysis of Asymmetric Hemoglobin Hybrids: Demonstration of Hb FS (α2γβS) in Sickle Cell Disease

Direct and indirect pathways of functional coupling in human hemoglobin are revealed by quantitative low-temperature isoelectric focusing of mutant hybrids

Metal Ion Reconstituted Hybrid Hemoglobins

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