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Measurement of Heme Synthesis Levels in Mammalian Cells
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Measurement of Heme Synthesis Levels in Mammalian Cells

Published on: July 9, 2015

Haem disorder in recombinant- and reticulocyte-derived haemoglobins: evidence for stereoselective haem insertion in

A J Mathews1, T Brittain

  • 1School of Biological Sciences, University of Auckland, 3 Symonds Street, Auckland, New Zealand.

The Biochemical Journal
|June 21, 2001
PubMed

We have used NMR spectroscopy to measure haem disorder in adult human haemoglobin (HbA) obtained from mature erythrocyte cells and from yeast expressing recombinant HbA. Reticulocyte-derived HbA contained much higher levels of haem disorder (11% alpha- and 28% beta-subunit disorder) than observed for HbA from mature erythrocytes (1.5% alpha- and 8% beta-subunit disorder). Thus, unlike in vitro combination of haem and apoHb, biosynthetic haem insertion is not random with respect to orientation, but appears to show stereoselectivity. Recombinant HbA isolated from yeast showed 32% alpha- and 45% beta-subunit haem disorder. These levels relaxed to their equilibrium positions after incubating the Hb in the ferric form. Recombinant embryonic human Hbs showed less haem disorder than recombinant HbA. The levels of haem disorder in embryonic Hbs zeta(2)epsilon(2) and zeta(2)gamma(2) appear to have their equilibrium values. We propose that, in eukaryotes, in vivo haem insertion occurs via both co-translational mechanisms and insertion via semiHb-beta.

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