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J. Biol. Chem., Vol. 275, Issue 51, 40187-40194, December 22, 2000

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Regulation of a Ca²⁺-sensitive Adenylyl Cyclase in an Excitable Cell
ROLE OF VOLTAGE-GATED VERSUS CAPACITATIVE Ca²⁺ ENTRY^*

Kent A. Fagan, Robert A. Graf, Shawna Tolman^§, Jerome Schaack^§, and Dermot M. F. Cooper^¶

From the Departments of  Pharmacology and ^§ Microbiology, University of Colorado Health Sciences Center, Denver, Colorado 80262

In nonexcitable cells, we had previously established that Ca²⁺-sensitive adenylyl cyclases, whether expressed endogenously or heterologously, were regulated exclusively by capacitative Ca²⁺ entry (Fagan, K. A., Mahey, R. and Cooper, D. M. F. (1996) J. Biol. Chem. 271, 12438-12444; Fagan, K. A., Mons, N., and Cooper, D. M. F. (1998) J. Biol. Chem. 273, 9297-9305). Relatively little is known about how these enzymes are regulated by Ca²⁺ in excitable cells, where they predominate. Furthermore, no effort has been made to determine whether the prominent voltage-gated Ca²⁺ entry, which typifies excitable cells, overwhelms the effect of any capacitative Ca²⁺ entry that may occur. In the present study, we placed the Ca²⁺-stimulable, adenylyl cyclase type VIII in an adenovirus vector to optimize its expression in the pituitary-derived GH₄C₁ cell line. In these cells, a modest degree of capacitative Ca²⁺ entry could be discerned in the face of a dramatic voltage-gated Ca²⁺ entry. Nevertheless, both modes of Ca²⁺ entry were equally efficacious at stimulating adenylyl cyclase. A striking release of Ca²⁺ from intracellular stores, triggered either by ionophore or thyrotrophin-releasing hormone, was incapable of stimulating the adenylyl cyclase. It thus appears as though the intimate colocalization of adenylyl cyclase with capacitative Ca²⁺ entry channels is an intrinsic property of these molecules, regardless of whether they are expressed in excitable or nonexcitable cells.

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