ISSN:
1573-4943
Keywords:
DPG
;
blood substitutes
;
α-globin
;
Β-chains
Source:
Springer Online Journal Archives 1860-2000
Topics:
Chemistry and Pharmacology
Notes:
Abstract Interspecies hybrids of HbA and Hb from mouse C57BL/10 [α 2 M Β 2 H and↠ 2 H Β 2 M (H=human, M=mouse)], representing 19 and 27 sequence differences perαΒ dimers (as compared with humanαΒ dimer) have been generatedin vitro. The efficiency of the assembly of the interspecies hybrids by the alloplex intermediate pathway is about twofold higher than the low-pH-mediated subunit approach. The interspecies hybrids exhibit a cooperative O2 binding. The intrinsic O2 affinity of mouse Hb is slightly lower than HbA, while the 2,3-diphosphoglycerate (DPG) effect is comparable. Interestingly, the interspecies hybridα 2 M Β 2 H has high O2 affinity (compared to either human or mouse Hb), while the interspecies hybridα 2 H Β 2 M exhibits a very low O2 affinity. These results suggest that the mouseΒ chain generates a tetramer with very low oxygen affinity. However, the complementarity of the mouseα andΒ chains generates a set of unique interactions that compensate for the low-oxygen-affinity propensity of the mouseΒ chain. DPG binds the tetramer in the central cavity formed by the twoΒ subunits, hence the DPG effects on the interspecies hybrids should be as in the parent molecule. However, the results of the present study demonstrate that the DPG binding pocket is influenced by the nature of theα chain present in the tetramer. The mouseα chain reduces considerably the DPG right shift of the O2 affinity of the humanΒ-chain containing hybrid. Sequence analysis suggest that perturbations of theα 1 Β 1 (not theα 1 Β 2) are communicated to the DPG binding pocket in the presence of the alien subunit, and are the primary determinant of the ligand binding properties. The results have implications for the design of Hb-based blood substitutes and understanding of the inhibitory potential of mouseα chains in transgenic mouse expressing humanΒ S chains.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01888365
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