G Protein-Coupled Receptors by Gabriel Berstein

By Gabriel Berstein

Overlaying lately constructed equipment in membrane-bound receptors, this ebook emphasizes receptor constitution and serve as, wisdom of that is necessary to the learn of sign transduction.
G Protein-Coupled Receptors has culled members from household and foreign resources, delivering a wide base of information. a few subject matters lined are the legislation of receptors; phosphorylation of G-protein-coupled receptors and ion channel receptors; desensitization reports utilizing electrophysiological or confocal microscopic tools and the quest for endogenous ligands for orphan receptors.
This quantity comes in handy at either the collage and commercial degrees, and is of specific curiosity to people who are constructing healing techniques to ailments utilizing medications that impact receptor activation.

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It is easy to detect depletion of the radioligand, since the concentrations of both added and bound material are measured routinely. The possibility of radioligand depletion should always be considered. 3,10,63 As discussed below, computational adjustments are unlikely to be completely accurate. If possible, it is preferable to avoid the problem by increasing the radioligand concentration or the incubation volume, or decreasing the tissue concentration, but practical limitations may sometimes make it impossible to prevent depletion, especially with very potent tritiated radioligands.

In the design of the assay, the concentration of radioligand to be used has to be carefully considered. The choice depends on the information required. On one hand it is important to know the time course of binding of the radioligand concentration used in other studies such as inhibition assays (see below), a concentration which is often around the KD level or below. 7 and below), provided the nonspecific binding is not too high. It is recommended that at least two radioligand concentrations be used.

If a logistic fit to data from an experiment has a slope different from 1, then the Cheng-Prusoff correction may be used to account for the competitive effect of the radioligand concentration, but the Ki value derived from such a fit is only an estimate of the IC50 value which would probably be obtained with a very low radioligand concentration, rather than an estimate of the intrinsic dissociation constant of the inhibitor. 5 ASSAY 5: ALLOSTERISM ANALYSIS The following assay is based on the allosteric model presented above (Scheme 4) and allows quantitative measurement of the interactions between an allosteric agent and both a radioligand and a second, unlabeled, primary ligand, under both equilibrium and nonequilibrium conditions.

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