Subtype-specific dimerization of alpha 1-adrenoceptors: effects on receptor expression and pharmacological properties.
The potential role of dimerization in controlling the expression and pharmacological properties of alpha1-adrenoceptor subtypes was examined using coimmunoprecipitation of epitope-tagged receptors. Human alpha1-adrenoceptor subtypes (alpha1A, alpha1B, alpha1D) were tagged at their amino-termini with Flag or hemagglutinin epitopes and transfected into human embryonic kidney 293 cells. Homodimerization of all three subtypes was observed by coimmunoprecipitation of receptors with different tags and was not altered by norepinephrine treatment. Heterodimer formation between hemagglutinin-tagged alpha1B-adrenoceptors and Flag-tagged alpha1A- or alpha1D-adrenoceptors was also observed. However, no alpha1A/alpha1D-adrenoceptor heterodimers were observed, suggesting that dimerization is subtype-specific. The extent of heterodimerization was also unaltered by norepinephrine treatment. alpha1-Adrenoceptor truncation mutants lacking carboxyl or amino-terminal sequences formed homo- and heterodimers similarly to full-length receptors, suggesting that these domains play little or no role in dimerization. Biotinylation with a membrane-impermeable agent showed that monomers and homo- and hetero-oligomers of all three subtypes are expressed on the cell surface. Radioligand binding studies showed that heterodimerization did not alter the affinity of alpha1-adrenoceptors for norepinephrine, prazosin, or subtype-selective antagonists, suggesting that dimerization does not result in pharmacologically distinct subtypes. However, coexpression of alpha1B-adrenoceptors significantly increased both binding site density and protein expression of alpha1A- and alpha1D-adrenoceptors, and increased cell surface expression of alpha1D-adrenoceptors, suggesting a functional role for heterodimerization. Conversely, coexpression of alpha1A-with alpha1D-adrenoceptors, which did not heterodimerize, had no effect on receptor density or protein. These studies demonstrate subtype-selective heterodimerization of alpha1-adrenoceptors, which does not change their pharmacological properties but seems to have functional consequences in regulating receptor expression and trafficking.
Uberti
Michelle A
MA
Department of Pharmacology, Emory University School of Medicine, 1510 Clifton Road, Atlanta, GA 30322, USA.
Hall
Randy A
RA
Minneman
Kenneth P
KP
D000317:Adrenergic alpha-Antagonists / Q000378:metabolism / Q000494:pharmacology; D002460:Cell Line; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D006801:Humans; D020033:Protein Isoforms / Q000096:biosynthesis / Q000145:classification / Q000378:metabolism / Q000502:physiology; D018340:Receptors, Adrenergic, alpha-1 / Q000096:biosynthesis / Q000145:classification* / Q000378:metabolism / Q000502:physiology*
D000317:Adrenergic alpha-Antagonists; D020033:Protein Isoforms; D018340:Receptors, Adrenergic, alpha-1