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. 1971 Sep;124(3):563–571. doi: 10.1042/bj1240563

Calcium binding by human erythrocyte membranes

Janet Forstner 1, J F Manery 1
PMCID: PMC1177226  PMID: 4332543

Abstract

1. The characteristics of Ca2+ binding to haemoglobin-free human erythrocyte membranes were investigated by using 45Ca and centrifugation partition of `ghosts' from their external incubation medium. Equilibrium of `ghosts' with external Ca2+ required less than 15min. 2. The binding did not vary with temperature in the range 0–37°C. 3. At pH7.4 `ghosts' bound a maximum of 283μmol of Ca2+/g of `ghost' protein, equivalent to 6.85×107 Ca2+ ions per cell. 4. Increasing the ionic strength from 0.01 to 0.46 diminished Ca2+ binding, as did ATP in concentrations ranging from 0 to 15mm in the incubation medium. 5. An increase of the pH from 3.0 to 9.3 caused a marked increase in the amount of Ca2+ bound. 6. Extraction of 45Ca-labelled `ghosts' with chloroform–methanol showed that the distribution of Ca2+ was: 79% protein-bound, 16% lipid-bound, 5% in the aqueous phase, presumably non-bound. Most of the lipid-bound Ca2+ (about 80%) was associated with a phospholipid fraction containing phosphatidylserine, phosphoinositides and phosphatidylethanolamine, giving a molar Ca2+: phosphorus ratio of about 1:2.

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Selected References

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