PMID: 3768348Sep 9, 1986Paper

1H NMR study of dynamics and thermodynamics of heme rotational disorder in native and reconstituted hemoglobin A

Biochemistry
Y Yamamoto, G N La Mar

Abstract

The reaction of heme and apoprotein has been studied in detail in 1H NMR spectroscopy in order to elucidate the conditions for reconstitution of hemoglobin (Hb) to yield the native protein. The initially formed holoprotein exists as a mixture of isomers with individual subunits possessing the two heme orientations differing by a 180 degrees rotation about the alpha, gamma-meso axis [La Mar, G. N., Yamamoto, Y., Jue, T., Smith, K. M., & Pandey, R. K. (1985) Biochemistry 24, 3826-3831]. We characterize in detail herein the rates and mechanism of heme reorientation and show that the rates differ dramatically for met-aquo and met-azido derivatives and are highly pH dependent in both subunits in a fashion that allows selective equilibration in either subunit. Nonequilibrium mixtures of such isomers can be kinetically trapped in the met-azido form and stored in this metastable form for many months. With kinetically controlled heme orientationally disordered Hb, unambiguous assignment of 1H NMR resonances to individual subunits has been made for the met-azido derivative, which demonstrates approximately 2% and 10% equilibrium heme disorder in the alpha- and beta-subunits, respectively. Comparison of the 1H NMR spectra of various heme ...Continue Reading

References

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Citations

Sep 20, 1991·Biochimica Et Biophysica Acta·J PaulK G Paul
Jan 31, 1989·Biochemical and Biophysical Research Communications·G N La MarM J Levy
Jul 4, 2002·Molecular Aspects of Medicine·Thomas Brittain
Feb 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·M L SmithK G Paul
Sep 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·T J ShenC Ho
Jun 13, 2002·Proceedings of the National Academy of Sciences of the United States of America·Oliver DaltropStuart J Ferguson
Apr 16, 1998·FEBS Letters·Y Yamamoto, T Nagaoka
Jan 16, 2014·Chirality·Masako NagaiSaburo Neya
Jan 30, 2007·Biochemical and Biophysical Research Communications·Satoshi NagaoYasuhiko Yamamoto
Oct 16, 2004·Biochemical and Biophysical Research Communications·Haruto IshikawaIsao Morishima
May 24, 2011·Biochimica Et Biophysica Acta·Sandrine JuillardGérard Simonneaux
Jun 28, 1988·Biochemistry·K Ishimori, I Morishima
Oct 1, 2011·The Journal of Physical Chemistry. B·Shigenori NagatomoHiroyuki Yoshikawa
Nov 4, 2004·Journal of the American Chemical Society·Teruyuki KomatsuEishun Tsuchida

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