Abstract
Parallel isofocusing studies established that carboxypeptidase A removal of the His-146 (HC3) and Tyr-145 (HC2) residues of β heme subunits affected the assembly properties of both Des β(A) and Des β(S) with α heme chains, albeit to differing degrees. Indeed, the rate of Des β(A) oligomer dissociation (k 1), as determined by visible spectroscopy, was 4.3-fold faster than that of its native β(A) counterpart. Furthermore, Soret spectral studies have affirmed distinct rates of normal (HbA), sickle (HbS), and Des HbA hemoglobin assembly (k′2) from their α and β [Des β(A)] heme-containing monomers. Matching kinetic analysis of Des β(A) and Des β(S) chain assembly (with an identical α chain) revealed 4.6- and 7.8-fold faster combination rates than those seen for β(A) and β(S) chains, respectively. This 3-fold disparity in rates strongly supports the critical role of the β-6 (A3) residue, and its amino-terminal region, in α chain partner recognition and subsequent human hemoglobin assembly.
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Morris, A., McDonald, M.J. Carboxyl-Terminal Modification Alters the Subunit Interactions and Assembly Pathways of Normal and Sickle Hemoglobins. J Protein Chem 20, 611–617 (2001). https://doi.org/10.1023/A:1013760017671
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DOI: https://doi.org/10.1023/A:1013760017671