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pubmed:abstractText |
Allosteric structure change in human hemoglobin was studied by hydrogen-tritium-exchange methods. The functional labeling method used takes advantage of the change in H-exchange rate at allosterically involved sites to selectively label, with tritium, H-exchange sites that are fast in one protein state and slow in another. The position of the labeled sites can then be located by the medium-resolution fragmentation-separation method. These methods reveal 5 allosterically sensitive, H-bonded, peptide NH's within the first 12 residues of the alpha chain. All five exchange with solvent protons at similar rates in deoxyhemoglobin (T form), and all shift to a new rate, about 30-fold faster, in the liganded protein (R) form. This indicates a decrease in structural stability at the alpha-chain N-terminus in going from the T to the R form, consistent with the loss of stabilizing interactions in that segment. The results indicate a loss of perhaps 2 kcal/mol in stabilization free energy and thus document a significant role for changes at the alpha-chain N-terminus in the allosteric transition.
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