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Pair-wise interactions of polymerization inhibitory contact site mutations of hemoglobin-S

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Abstract

The linkage of pair-wise interactions of contact site mutations of HbS has been studied using Le Lamentin [His-20 (α)→Gln], Hoshida [Glu-43 (β)→Gln] and α2β T87Q2 mutations as the prototype of three distinct classes of contact sites of deoxy HbS fiber. Binary mixture experiments established that βA-chain with the Thr-87 (β)→Gln mutation is as potent as the γ-chain of HbF (α2γ2) in inhibiting polymerization. On combining the influence of Le Lamentin mutation with that of β T87Q2 mutations; the net influence is only partial additivity. On the other hand, in binary mixture studies, combined influence of Hoshida mutation with that of β T87Q2 mutations is synergistic. Besides, a significant level of synergistic complementation is also seen when the Le Lamentin and Hoshida mutations are combined in HbS (symmetrical tetramers). Le Lamentin and Hoshida mutation introduced into the cis-dimer of the asymmetric hybrid tetramer completely neutralizes the Val-6 (β) dependent polymerization. Accordingly, we propose that combining the perturbation of intra-double strand contact site with that of an inter-double strand contact site exhibit synergy when they are present in two different chains of the αβ dimer. A comparison of the present results with that of the earlier studies suggest that when the two contact site perturbations are from the same sub-unit of the αβ dimer only partial additivity is observed. The map of interaction linkage of the contact site mutations exposes new strategies in the design of novel anti-sickling Hbs for the gene therapy of sickle cell disease.

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Abbreviations

HbA:

Adult human hemoglobin

HbS:

Sickle Cell hemoglobin

HbF:

Fetal hemoglobin

SCA:

Sickle cell anemia

WL-DS:

Wishner Love double strand

TFA:

Trifluro acetic acid

HPLC:

High performance liquid chromatography

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Acknowledgments

This work was supported by following National Institute of Health grants; NIH/NHLBI 1M01 RR 12248, NIH/NHLBI 5P60HL-38655, NIH/NHLBI P01HL55435, and PO1 HL58512.

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Correspondence to A. Seetharama Acharya.

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Srinivasulu, S., Perumalsamy, K., Upadhya, R. et al. Pair-wise interactions of polymerization inhibitory contact site mutations of hemoglobin-S. Protein J 25, 503–516 (2006). https://doi.org/10.1007/s10930-006-9034-3

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