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α1-Adrenergic Receptor Signaling via Gh Is Subtype Specific and Independent of Its Transglutaminase Activity
Journal article   Open access   Peer reviewed

α1-Adrenergic Receptor Signaling via Gh Is Subtype Specific and Independent of Its Transglutaminase Activity

Songhai Chen, Fang Lin, Siiri Iismaa, Kyung N Lee, Paul J Birckbichler and Robert M Graham
The Journal of biological chemistry, Vol.271(50), pp.32385-32391
12/13/1996
DOI: 10.1074/jbc.271.50.32385
url
https://doi.org/10.1074/jbc.271.50.32385View
Published (Version of record) Open Access

Abstract

Tissue transglutaminase (TGase II) is a Ca2+- and thiol-dependent enzyme that catalyzes the post-translational modification of proteins via the formation of ϵ(γ-glutamyl) lysine bonds. We have shown previously that the GTP-binding protein, Gh, is a TGase II that mediates intracellular signaling by the α1B-adrenergic receptor (AR) (Nakaoka, H., Perez, D. M., Baek, K. J., Das, T., Husain, A., Mison, K., Im, M.-J., and Graham, R. M. (1994) Science 264, 1593-1596). Here, we evaluated the ability of Gh as compared with Gq to mediate receptor-stimulated inositol phosphate turnover by the three α1-subtypes (α1A, α1B, and α1D). In addition, we questioned if the transglutaminase function of Gh is involved in its receptor signaling activity. A mutant form of a human TGase II cDNA in which the codon for the active site cysteine (Cys277) was replaced by serine was cloned into the mammalian expression vector pMT2′. Compared with wild-type TGase II, no transglutaminase activity was observed with transient transfection of this Cys→Ser mutant in COS-1 cells. However, like wild-type TGase, the Cys→Ser mutant mediated receptor-stimulated inositol phosphate turnover when cotransfected with an α1B-AR cDNA. Gαq supported α1-AR-mediated inositol phosphate turnover by all three receptor subtypes. By contrast, although both the wild-type and Cys→Ser construct mediated receptor signaling by the α1B AR and α1D AR, the α1A-AR was unable to interact with Gh. However, a Gh-dependent signaling phenotype could be rescued by a chimeric α1A construct in which the third intracellular loop of the α1A-AR was replaced by that of the α1B-AR. Thus, the signaling function of Gh is independent of its transglutaminase activity and is α1-AR subtype specific. This subtype specificity of the interaction between α1 ARs and Gh involves important determinants in their third intracellular loops.

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