Inhibitory effect of isoproterenol on NADPH-dependent H(2)O(2) generation in human adipocyte plasma membranes is mediated by betagamma-subunits derived from G(s).
Previous studies revealed that human fat cell plasma membranes contain a multireceptor-linked H(2)O(2)-generating system that is under antagonistic control by hormones and cytokines and is stimulated by insulin via Galpha(i2). In this report, it is shown that the inhibitory action of the beta-adrenergic agonist isoproterenol is mediated by G protein betagamma-subunits, based on observations that its action was specifically reversed by anti-Gbeta antibodies or a C-terminal beta-adrenergic receptor kinase-1 fragment containing the Gbetagamma-binding site of the enzyme, and was mimicked by exogenously supplied G protein betagamma-subunits. Isoproterenol signals through a prototypical G(s)-coupled receptor. Consistent with these results, direct activation of G(s) by cholera toxin or by an anti-Galpha(s) antibody exhibiting beta-adrenergic receptor-mimetic properties (K-20) resulted in an isoproterenol-like inhibition of NADPH-dependent H(2)O(2) generation. In addition, a peptide corresponding to the target sequence of K-20 blocked the action of the catecholamine, apparently by competition between the peptide and G(s) for activated beta-adrenergic receptors, indicating that the G protein betagamma-subunits mediating the inhibitory effects of the catecholamine were in fact derived from G(s).
Krieger-Brauer
H I
HI
Klinisches Institut für Herzinfarktforschung an der Medizinischen Universitätsklinik Heidelberg, Bergheimerstrasse 58, D-69115 Heidelberg, Germany.
Medda
P K
PK
Sattel
B
B
Kather
H
H
D017667:Adipocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D007545:Isoproterenol / Q000494:pharmacology*; D008969:Molecular Sequence Data; D019255:NADPH Oxidases / Q000378:metabolism
D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006861:Hydrogen Peroxide; D019255:NADPH Oxidases; D019205:GTP-Binding Protein alpha Subunits, Gs; D007545:Isoproterenol