Abstract
The specific binding of Leu-enkephalin and the stimulatory effect of the peptide on cAMP accumulation have been assessed in isolated enterocytes of guinea pig. The binding was reversible as well as time and temperature dependent. Two classes of binding sites could be defined: a class with a relatively high affinity (K d=0.7 μM) that represented 1% of total binding capacity, and another class with low affinity (K d=55.5 μM). The stimulation of cAMP accumulation was also shown to depend on time and temperature and was potentiated by a phosphodiesterase inhibitor. Half-maximal stimulation of cAMP accumulation was observed at 119 μM and maximal stimulation (27-fold basal level) at 300 μM Leu-enkephalin. Both steps of the interaction were not modified by Na + but exhibited a high specificity since modifications in the structure of Leu-enkephalin resulted in an important loss of binding affinity and stimulatory activity.
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