Abstract
The expression of β-globin in Escherichia coli has enabled us to study the functional role of individual amino-acid residues in haemoglobin (Hb) by site-directed mutagenesis1. In contrast to mammalian Hbs, some teleost fish haemoglobins show a drastic lowering of oxygen affinity and cooperativity at low pH, a phenomenon known as the Root effect2. We have produced the two mutant haemoglobins Hb Nymphéas [Cys(F9)93β →Ser] and Hb Daphne [His(H21)143β → Arg, Cys(F9)93β → Ser] to investigate this allosteric property. Although these substitutions were thought to be responsible for the Root effect3, Hb Nymphéas and Hb Daphne show an increased oxygen affinity and a reduced effect of pH on oxygen affinity1. Our X-ray Crystallographic studies show that the hydroxyl group of Ser 93β forms a hydrogen bond with Asp 94β which is in equilibrium with the salt bridge between Asp 94β and His 146β. The oxygen-binding properties of Hbs Nymphéas and Daphne are accounted for by the partial disruption of the salt bridge.
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Luisi, B., Nagai, K. Crystallographic analysis of mutant human haemoglobins made in Escherichia coli. Nature 320, 555–556 (1986). https://doi.org/10.1038/320555a0
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DOI: https://doi.org/10.1038/320555a0
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