Comparative expression of the human beta(2) and beta(3) adrenergic receptors in Saccharomyces cerevisiae.
The beta(3) adrenergic receptor (beta(3)AR) is the predominant beta subtype in human brown adipocytes and is essential for regulating thermogenic lipolysis. To establish a novel experimental system for the biochemical analysis of this protein, we engineered several yeast strains. We show that the sterol background of the host strain greatly modulates the beta(3)AR expression but not in the same way as it modulates the beta(2) adrenergic receptor (beta(2)AR), the other main studied adipocyte subtype. The human beta(3)AR expressed in yeast is N-glycosylated but not phosphorylated. This latter characteristic distinguishes it from the beta(2)AR. We showed that both beta(2)AR and beta(3)AR follow the secretory pathway to the yeast plasma membrane (PM) and are degraded in the vacuole. In the yeast strains used in this work, the two receptors also share a common mechanism of direct signal transduction through the yeast G(alpha) protein, Gpa1p. These strains thus appear to be useful for biochemical and structural studies of the human beta(3)AR in an in vivo reconstitution system.
Duport
Catherine
C
Université Paris VII et Institut Cochin de Génétique Moléculaire, 22 rue Méchain, 75014 Paris, France. duport@avignon.inra.fr
Loeper
Jacqueline
J
Strosberg
A Donny
AD
D002462:Cell Membrane / Q000737:chemistry; D044385:GTP-Binding Protein alpha Subunits; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D015870:Gene Expression; D020962:Heterotrimeric GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D011485:Protein Binding; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D022702:Receptors, Adrenergic, beta-3 / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D015398:Signal Transduction; D013261:Sterols / Q000378:metabolism; D014162:Transfection
D044385:GTP-Binding Protein alpha Subunits; D018343:Receptors, Adrenergic, beta-2; D022702:Receptors, Adrenergic, beta-3; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D013261:Sterols; C084293:GPA1 protein, S cerevisiae; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020962:Heterotrimeric GTP-Binding Proteins