A mutagenic study of the allosteric linkage of His(HC3)146 beta in haemoglobin
| Autor: | Daniel T.-b. Shih, Gentaro Miyazaki, Ben F. Luisi, Max F. Perutz, Kiyoshi Nagai |
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| Rok vydání: | 1993 |
| Předmět: |
Hemeprotein
Magnetic Resonance Spectroscopy Stereochemistry Protein Conformation Hemoglobins Abnormal Allosteric regulation Bohr effect Sodium Chloride Protein structure Allosteric Regulation X-Ray Diffraction Structural Biology Escherichia coli Humans Histidine Beta (finance) Molecular Biology Hydrogen bond Chemistry Mutagenesis Hemoglobin A Hydrogen Bonding Hydrogen-Ion Concentration Recombinant Proteins Oxygen Kinetics Models Chemical Mutagenesis Site-Directed Hemoglobin |
| Zdroj: | Journal of molecular biology. 230(4) |
| ISSN: | 0022-2836 |
| Popis: | We have examined the contribution of His(HC3)146 beta to the alkaline Bohr effect of human haemoglobin (HbA) by replacing it with Gln, using site-directed mutagenesis, and studying the structural and functional consequences. Oxygen equilibrium curves of the mutant show that the effect of pH on the oxygen affinity, the alkaline Bohr effect, is half that of HbA in the presence of chloride ion and less than 10% in its absence. Crystallographic analysis shows that the mutation introduced only small structural changes localized to the site of substitution, proving that the replacement of the hydrogen bond between the ionizable side-chain of His146 beta and Asp94 beta by a hydrogen bond between the unionizable side-chain of Gln146 beta and the same aspartate is solely responsible for the reduction of the alkaline Bohr effect. Our data confirm that His(HC3)146 beta is predominantly responsible for the chloride-independent component of the alkaline Bohr effect which results from the breaking of the hydrogen bond between His(HC3)146 beta and Asp(FG1)94 beta accompanying the transition from the quaternary deoxy to oxy-structure. |
| Databáze: | OpenAIRE |
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