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Stabilizing and destabilizing effects on plasma membrane Ca(2+)-ATPase activity

D Kosk-Kosicka1, A Wawrzynow, G Roszczynska

  • 1Department of Anesthesiology/CCM, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287.

Molecular and Cellular Biochemistry
|October 12, 1994
PubMed
Summary

Polyols stabilize erythrocyte Ca(2+)-ATPase activity across temperatures, unlike other organic compounds. Glycerol offers protection against thermal denaturation, while other solutes inhibit the enzyme through destabilizing interactions.

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Area of Science:

  • Biochemistry
  • Enzymology
  • Membrane Proteins

Background:

  • The Ca(2+)-ATPase enzyme is crucial for calcium ion transport in erythrocytes.
  • Enzyme activity is modulated by temperature and interactions with various organic compounds.
  • The enzyme exists in monomeric (calmodulin-activated) and oligomeric forms.

Purpose of the Study:

  • To investigate the temperature-dependent effects of organic compounds on purified erythrocyte Ca(2+)-ATPase activity.
  • To differentiate between stabilizing and destabilizing interactions of solutes with the enzyme.
  • To compare the effects of solutes on monomeric and oligomeric forms of the enzyme.

Main Methods:

  • Purification of Ca(2+)-ATPase from erythrocytes.
  • Assay of enzyme activity at varying temperatures and solute concentrations.

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  • Analysis of homologous series of organic compounds: polyols, alkanols, aprotic solvents, and N-methyl derivatives.

    • Main Results:

    • Polyols demonstrated a broad stabilizing effect on Ca(2+)-ATPase activity across concentration and temperature ranges.
    • Glycerol, a polyol, protected the enzyme from thermal denaturation at 45°C.
    • Other solutes (alkanols, solvents, amides) exhibited inhibitory effects, increasing with concentration and structural similarity to detergents, comparable to urea.
    • Oligomeric Ca(2+)-ATPase showed greater resistance to inhibitory solutes than calmodulin-activated monomers.

    Conclusions:

    • Polyols provide indirect, stabilizing interactions, maintaining enzyme activity at different temperatures.
    • Direct, destabilizing interactions are responsible for the inhibition observed with other organic compounds.
    • The oligomeric structure of Ca(2+)-ATPase offers enhanced protection against solute-induced destabilization.