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X Zhu, S Gilbert, M Birnbaumer and L Birnbaumer
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
We studied the relative ability to activate phospholipase C (PLC) of four Gs-coupled receptors expressed in L cells at different densities. Stable cell lines expressing various levels of the luteinizing hormone receptor (LHR), the type 2 vasopressin receptor (V2R), or the type 1 or type 2 beta-adrenergic receptor (beta 1- or beta 2AR) were isolated. The PLC activity was assessed by the measurement of free intracellular Ca2+ concentrations and the accumulation of inositol phosphates. We previously reported that, at 24,000 sites/cell, the LHR in L cells stimulated adenylyl cyclase by 10-fold over basal levels and PLC by 50% over basal levels. The EC50 for stimulation was 20-fold higher for PLC than for adenylyl cyclase. We now report that LHR tends to stimulate PLC more at a higher receptor density and less at a lower density. EC50 values for accumulation of inositol phosphates remained unchanged. The human V2R and the human beta ARs are strong adenylyl cyclase stimulators, and their potential for dual signaling was unknown. Expressing the V2R at 100,000 sites/cell or more and the beta ARs at 300,000 sites/cell resulted in stimulation of PLC by these receptors. As with the LHR, higher concentrations of vasopressin or isoproterenol were needed to reach 50% stimulation of PLC, compared with that of adenylyl cyclase. The beta 1AR was a stronger PLC stimulator than was the beta 2AR. The orders of potency for isoproterenol, epinephrine, and norepinephrine to stimulate adenylyl cyclase and PLC were the same for each of the two beta ARs. These results indicate that the ability of Gs- coupled receptors to stimulate PLC is dependent on the levels of receptor expression, and they suggest that dual signaling potential is a common property of Gs-coupled receptors and possibly also of G(i)- coupled receptors.
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