Sub cellular Fractionation of the Longitudinal Smooth Muscle/Myenteric Plexus of Dog Ileum: Dissociation of the Distribution of Two Plasma Membrane Marker Enzymes

Abstract

The distribution of plasma membrane markers, the sodium pump [evaluated as ouabain-sensitive, potassium-stimulated p-nitrophenyl phosphatase (K⁺-pNPPase)], [³H]saxitoxin binding, and 5′-AMPase, was studied in the subcellular fractions prepared from the ho-mogenates of the longitudinal smooth muscle/myenteric plexus of dog ileum. The K⁺-pNPPase activity and [³H]-saxitoxin binding were found to be predominantly associated with the synaptosomal fraction as indicated by the high level of these activities in the crude synaptosomal fraction and by the copurification of K⁺-pNPPase and [³H]saxitoxin binding, but not 5′-AMPase, with several synaptosomal markers during the fractionation of the crude synaptosomal fraction on density gradients. In contrast to the K⁺-pNPPase activity and [³H]saxitoxin binding, the 5′-AMPase activity was found to be concentrated in the microsomal pellet. Further fractionation of microsomes on a density gradient resulted in copurification of 5′-AMPase, but not K⁺-pNPPase or [³H]saxitoxin binding, with other smooth muscle plasma membrane-bound enzymes, such as high-affinity Ca²⁺-ATPase, Mg²⁺-ATPase, and Ca²⁺-ATP-ase. It was concluded that in the longitudinal smooth muscle/myenteric plexus, the sodium pump activity is present in higher density in the neuronal plasma membranes whereas 5′-AMPase activity is concentrated in the smooth muscle plasma membranes.