Semihemoglobins, high oxygen affinity dimeric forms of human hemoglobin respond efficiently to allosteric effectors without forming tetramers

Antonio Tsuneshige, Kenji Kanaori, Uri Samuni, David Danstker, Joel M. Friedman, Saburo Neya, Laura Giangiacomo, Takashi Yonetani

Research output: Contribution to journalArticle

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Abstract

Significant reduction in oxygen affinity resulting from interactions between heterotropic allosteric effectors and hemoglobin in not only the unligated derivative but also the fully ligated form has been reported (Tsuneshige, A., Park, S. I., and Yonetani, T. (2002) Biophys. Chem. 98, 49-63; Yonetani, T., Park, S. I., Tsuneshige, A., Imai, K., and Kanaori, K. (2002) J. Biol. Chem. 277, 34508-34520). To further investigate this effect in more detail, α- and β-semihemoglobins, namely, α(heme)β(apo) and α(apo)β(heme), respectively, were prepared and characterized with respect to the impact of allosteric effectors on both conformation and ligand binding properties. Semihemoglobins are dinners characterized by a high affinity for oxygen and lack of cooperativity. We found that, compared with stripped conditions, semihemoglobins responded to effectors (inositol hexaphosphate and L35) by decreasing the affinity for oxygen by 60- and 130-fold for α- and β-semihemoglobins, respectively. 1H NMR and sedimentation velocity experiments carried out with their ligated and unligated forms in the absence and presence of effectors revealed that semihemoglobins always remain as single-heme-carrying dimers. Recombination kinetics of their photolyzed CO derivatives showed that effectors did indeed interact with their ligated forms. Measurements of the Fe-His stretching mode snow that the semihemoglobins undergo a large ligand binding-induced conformational shift and that both ligand-free and ligand derivatives respond to the presence of effectors. Contradictions to the Monod-Wyman-Changeaux/Perotz allosteric model arise since 1) the modulation of ligand affinity is not achieved in semihemoglobins by the formation of a low affinity T conformation (quaternary effect) but by direct interaction with effectors, 2) effectors do interact significantly with ligated forms of high affinity semihemoglobins, and 3) modulation of the ligand affinity and the cooperativity are not necessarily linked but instead can be separated into two distinct phenomena that can be isolated.

Original languageEnglish (US)
Pages (from-to)48959-48967
Number of pages9
JournalJournal of Biological Chemistry
Volume279
Issue number47
DOIs
StatePublished - Nov 19 2004

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Hemoglobins
Oxygen
Ligands
Heme
Derivatives
L 35
Conformations
Modulation
Snow
Phytic Acid
Carbon Monoxide
Sedimentation
Dimers
Genetic Recombination
Stretching
Meals
Nuclear magnetic resonance
Kinetics
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Semihemoglobins, high oxygen affinity dimeric forms of human hemoglobin respond efficiently to allosteric effectors without forming tetramers. / Tsuneshige, Antonio; Kanaori, Kenji; Samuni, Uri; Danstker, David; Friedman, Joel M.; Neya, Saburo; Giangiacomo, Laura; Yonetani, Takashi.

In: Journal of Biological Chemistry, Vol. 279, No. 47, 19.11.2004, p. 48959-48967.

Research output: Contribution to journalArticle

Tsuneshige, Antonio ; Kanaori, Kenji ; Samuni, Uri ; Danstker, David ; Friedman, Joel M. ; Neya, Saburo ; Giangiacomo, Laura ; Yonetani, Takashi. / Semihemoglobins, high oxygen affinity dimeric forms of human hemoglobin respond efficiently to allosteric effectors without forming tetramers. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 47. pp. 48959-48967.
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