A Mutant (Arg³²⁷→His) GPIIb Associated to Thrombasthenia Exerts a Dominant Negative Effect in Stably Transfected CHO Cells

Summary

This work reports the structural and functional characterization of the platelet glycoprotein complex GPIIb-IIIa (integrin α_IIbβ₃) in a patient of type II Glanzmann thrombasthenia, bearing a homozygous G→A base transition at position 1074 of GPIIb that results in an Arg³²⁷→His substitution.

CHO cells stably transfected with cDNA encoding His³²⁷GPIIb showed a drastic reduction in the surface expression of α_IIbβ₃ complex relative to control cells transfected with wild type GPIIb. Immunopre-cipitation analysis demonstrated that GPIIb synthesis, heterodimeriza-tion, and short term maturation were not impeded, suggesting that conformational changes dependent on Arg³²⁷ of GPIIb may play an essential role in either the rate of maturation and/or transport of heterodimers to the cell surface.

Cotransfection of CHO cells with equimolar amounts of cDNAs encoding wild type and mutant His³²⁷-GPIIb led to a marked reduction in the surface expression of α_IIbβ₃. This novel observation of a dominant-negative effect of the mutant His³²⁷α_IIb subunit provides a molecular basis for the reduced platelet α_IIbβ₃ content observed in the heterozygous offspring.

• References

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