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Using 125I-labeled neurotensin (NT), porcine brain membranes were found to contain two types of high-affinity receptors, one class (approximately 1/3 of total) with an apparent Kd of 0.12 nM and another with an apparent Kd of 1.4 nM. Nonhydrolyzable analogs of GTP inhibited NT binding in a dose-dependent manner. In the presence of 60 microM guanosine(More)
An increased turnover of phosphatidate and phosphatidyl inositol has been found in many tissues where hormones or neurotransmitters are postulated to raise Ca2+ influx, for example in smooth muscle. However, the relationship between changes in phospholipid metabolism and changes in Ca2+ permeability was unknown. Following recent reports on the interactions(More)
The mechanism of beta-adrenergic relaxation was investigated in isolated smooth muscle cells. Beta-adrenergic agents stimulate cyclic AMP-dependent phosphorylation, enhance Na+/K+ transport and induce relaxation. The stimulation of Na+/K+ transport is obligatory for relaxation, and we suggest that this stimulation induces relaxation through enhanced(More)
Ligand binding properties of muscarinic receptors were examined in membranes and isolated cells prepared from bovine trachea. The binding of the muscarinic antagonist [3H]quinuclidinyl benzilate (QNB) to both membranes and isolated cells was saturable, reversible, and of high affinity [dissociation constant (KD) = 100-200 pM]. The binding constants of three(More)
The Ca2+-selective, fluorescent dye, fura-2, was used to monitor ATP-dependent Ca2+ uptake by membrane vesicles isolated from rabbit skeletal muscle. Micromolar fura-2 concentrations, added outside the vesicles, served as a high-affinity, low-capacity Ca2+ buffer. Changes in fura-2 fluorescence resulted from the decline in free Ca2+ concentration [((More)
The interaction of cholinergic agonists and antagonists with smooth muscle muscarinic receptors has been investigated by measurement of displacement of the muscarinic antagonist [3H]QNB (quinuclidinyl benzilate) in membranes prepared from toad stomach. The binding of [3H]QNB was saturable, reversible and of high affinity (KD = 423 pM). The muscarinic(More)
Studies were carried out to determine the effects of the beta-adrenergic agent, isoproterenol (ISO), on membrane electrical properties in single smooth muscle cells enzymatically dispersed from toad stomach. In cells bathed in buffer of physiological composition, the average resting potential was -56.4 +/- 1.4 mV (mean +/- SE, n = 35). The dominant effect(More)
The relationship between human erythrocyte glucose transporter (GLUT1) oligomeric strctUe and function was studied. GLUTI was purfied from human erythocytes in the absence (GLUTI-DTT) or the presence (GLUT I +DTT) of dithiothreitol. Chemical cross-linking studies of lipid bilayer-resident purfied GLUTI and hydrodynamic studies of cholate-solubilzed GLUTI(More)
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