Erythrocyte membrane thiol proteins associated with changes in the kinetics of Na/Li countertransport: a possible molecular explanation of changes in disease

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Abstract

Background

Abnormal erythrocyte Na/Li countertransport is associated with diseases such as essential hypertension and diabetic renal disease. Although it seems unlikely that Na/Li countertransport contributes to any disease process, it may be abnormal because of a change in the cell membrane that is part of the disease process.

Methods

We have shown that Na/Li countertransport kinetics are modified by two types of thiol group. One of these, which we have called 'type 1', is rapidly alkylated by N-ethylmaleimide to give a kinetic pattern similar to that in the above diseases.

Results

At pH 6 and 2 °C, both N-ethylmaleimide and iodoacetamide cause the Km of Na/Li countertransport to decrease to completion in 300 s, with 78% (SEM 6%) of the decrease occurring in 30 s. Using these reaction conditions, N-ethylmaleimide reacted with a unique thiol group on a 33-kD protein, blocking its subsequent reaction with biotin maleimide. This 33-kD protein was present in rabbit erythrocytes, which have high levels of Na/Li countertransport, but absent from rat erythrocytes, which have no Na/Li countertransport. Iodoacetyl biotin labelled a 60-kD protein that was specifically blocked by iodoacetamide.

Conclusion

We suggest that these proteins are members of a cluster of membrane proteins that can modify Na/Li countertransport and may have a functional role in the disease processes.

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