Solubilization and characterization of functionally coupled Escherichia coli heat-stable toxin receptors and particulate guanylate cyclase associated with the cytoskeleton compartment of intestinal membranes

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Abstract

  • 1.

    1. Particulate guanylate cyclase and receptors for E. coli heat-stable enterotoxin were solubilized from the rat intestinal cytoskeletal compartment using Lubrol-PX and KC1.

  • 2.

    2. Thirty to forty percent of the ST receptor and guanylate cyclase activities were extracted from the lipid layer with Lubrol-PX alone.

  • 3.

    2. Seventy percent of the remaining activities were solubilized from the cytoskeleton with Lubrol-PX and KCl.

  • 4.

    3. Guanylate cyclase solubilized from either compartment exhibited similar reaction kinetics.

  • 5.

    4. Both high- and low-affinity classes of ST receptors were solubilized from the lipid and cytoskeleton compartments.

  • 6.

    5. In the presence of ATPγS, ST selectively activated the guanylate cyclase solubilized from the cytoskeleton compared to that solubilized from the lipid bilayer.

  • 7.

    6. Crosslinking experiments demonstrated a preferential solubilization of the 130 kDa receptor subunit from the cytoskeleton and the 56 kDa subunit from the lipid bilayer.

  • 8.

    7. Development of a procedure to solubilize ST receptors and guanylate cyclase from the intestinal membrane cytoskeleton will permit purification and further detailed studies of the coupling of these activities.

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