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Noradrenaline induces binding of Clathrin light chain A to α1-adrenoceptors in the human prostate

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Abstract

BACKGROUND

Binding of clathrins or caveolin to G protein-coupled receptors may induce post-translational modifications of receptor function. Receptor regulation by clathrin requires cofactors ADP-ribosylation factor 6 (ARF6) and adaptin, while dynamin is required for clathrin- and caveolin-dependent mechanisms.

OBJECTIVE

To investigate the expression and α1-adrenoceptor binding of clathrins, caveolin, and their cofactors in the human prostate.

METHODS

Prostate tissue was obtained from radical prostatectomy. Expression of clathrin heavy chain (HC), clathrin light chain A and B (LCA, LCB), caveolin-1, ARF6, β-adaptin, and dynamin-2 was studied by RT-PCR, Western blot, immunohistochemistry, and fluorescence staining. Interaction of α1A-adrenoceptors with clathrins and caveolin-1 was studied by coimmunoprecipitation.

RESULTS

mRNA and protein expression of clathrin HC, LCA, LCB, caveolin-1, dynamin-2, and β-adaptin was detected in prostate tissues of each patient. Immunohistochemistry demonstrated the expression of clathrin HC, LCA, LCB, caveolin, dynamin, and β-adaptin in stromal cells. Immunoreactivity for these proteins colocalized with α-smooth muscle actin and α1A-adrenoceptors in double fluorescence staining. Coimmunoprecipitation demonstrated that α1A-adrenoceptors in prostate tissue interact with clathrin HC and LCB under resting conditions, but not with caveolin-1. Stimulation of prostate tissues with noradrenaline (30 µM) in vitro induced binding of clathrin LCA to α1A-adrenoceptors.

CONCLUSIONS

The prostatic α1-adrenoceptor population is at least partially bound to clathrin HC and LCB. Upon receptor activation, prostate α1A-adrenoceptors bind clathrin LCA. This points to a new concept of post-translational α1-adrenoceptor regulation in the prostate, which includes receptor interaction with accessory binding partners. Prostate 73: 715–723, 2013. © 2013 Wiley Periodicals, Inc.

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