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Biophysical Journal 86:2251-2260 (2004)
© 2004 The Biophysical Society

Divalent Cations Increase Lipid Order in Erythrocytes and Susceptibility to Secretory Phospholipase A2

Rebekah S. Vest *, Laurie J. Gonzales *, Seth A. Permann *, Emily Spencer *, Lee D. Hansen {dagger}, Allan M. Judd * and John D. Bell *

* Department of Physiology and Developmental Biology, and {dagger} Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah

Correspondence: Address reprint requests to John D. Bell, Dept. of Physiology and Developmental Biology, Brigham Young University, Provo, UT 84602. Tel.: 801-422-2353; Fax: 801-422-0050; E-mail: john_bell{at}byu.edu.

Elevated concentrations of intracellular calcium in erythrocytes increase membrane order and susceptibility to secretory phospholipase A2. We hypothesize that calcium aids the formation of domains of ordered lipids within erythrocyte membranes by interacting directly with the inner leaflet of the cell membrane. The interface of these domains with regions of more fluid lipids may create an environment with weakened neighbor-neighbor interactions that would facilitate phospholipid migration into the active site of bound secretory phospholipase A2. This hypothesis was investigated by determining the effects of seven other divalent ions on erythrocyte membrane properties. Changes in membrane order were assessed with steady-state fluorescence spectroscopy and two-photon microscopy with an environment-sensitive probe, laurdan. Each ion increased apparent membrane order in model membranes and in erythrocytes when introduced with an ionophore, suggesting that direct binding to the inner face of the membrane accounts for the effects of calcium on membrane fluidity. Furthermore, the degree to which ions affected membrane properties correlated with the ionic radius and electronegativity of the ions. Lastly, erythrocytes became more susceptible to enzyme hydrolysis in the presence of elevated intracellular levels of nickel and manganese, but not magnesium. These differences appeared related to the ability of the ions to induce a transition in erythrocyte shape.




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