G-protein Mechanism

Sprang, S.R. G protein mechanisms insights from structural analysis. Annu. Rev. Biochem. 66 639-678, 1997. 

FIGURE 5.3 Different types of functional readouts of agonism. Receptors need not mediate cellular response but may demonstrate behaviors such as internalization into the cytoplasm of the cell (mechanism 1). Receptors can also interact with membrane proteins such as G-proteins (mechanism 2) and produce cytosolic messenger molecules (mechanism 3), which can go on to mediate gene expression (mechanism 4). Receptors can also mediate changes in cellular metabolism (mechanism 5). 

Anandamide has dual effects on NMDA receptor function. It reduces NMDA Ca2-i- currents, which is mediated by cannabinoid receptors and G-protein mechanisms (Hampson et al. 1998). However, anandamide potentiates NMDA currents due to a direct effect on the NMDA receptor itself. THC did not have the same effect. So anandamide appears to have at least one other CNS effect that is not through the cannabinoid receptors. THC also reduces AMPA/kainate activity, which is mediated by CBl receptors (Shen and Thayer 1999) (table 10.5). THC may in-... 

Neubig, R. R. (1994). Membrane organization in G-protein mechanisms. FASEB J. 8, 939-946. 

Since the orexin receptors are Gq protein-coupled (Alexander et al. 2006), one may assume that this also holds true for the presynaptic orexin receptor(s), but so far no data are available. Nonetheless, the six studies carried out in central nervous preparations permit some conclusions on the post-G protein mechanisms. In all instances, the orexins increased the frequency of spontaneous inhibitory or excitatory postsynaptic potentials or currents. The results differed, however, with respect to the influence of tetrodotoxin. In the medial and lateral hypothalamus (van den Pol et al. 1998 Li et al. 2002), dorsal vagal complex (Davis et al. 2003), and caudal nucleus tractus solitarii (Smith et al. 2002), orexins increased the frequency of the miniature potentials or currents also in the presence of tetrodotoxin, suggesting that they directly influenced the vesicle release machinery (references in italics in Table 5). On the other hand, in the prefrontal cortex (Lambe and Aghajanian 2003) and lat-erodorsal tegmentum (Burlet et al. 2002), the orexins did not retain their facilitatory effect in the presence of tetrodotoxin, suggesting an effect further upstream e.g., on Ca2+ and/or K+ channels. 

Siderovski DP, Willard FS. The GAPs, GEFs, and GDIs of het-erotrimeric G-protein alpha subunits. Int. J. Biol. Sci. 2005 1 51-66. Sprang SR. G protein mechanisms insights from structural analysis. Ann. Rev. Biochem. 1997 66 639-678. 

Overholt JL, Prahhakar NR. Norepinephrine inhibits a toxin resistant Ca current in carotid body glomus cells evidence for a direct G protein mechanism. J Neurophysiol 1999 81 225-233. 

Some G proteins are slow GTP hydrolases with turnover numbers around two per minute, others such as Ras are only marginally catalytic. Kinetic experiments in solution have shown that in both cases the most likely mechanism... 

Sondek, J., et al. GTPase mechanism of G proteins from the 1.7 A crystal structure of transducin a.GDP.AlF4 . Nature 372 276-279, 1994. 

The most probable mechanism for inverse agonism is the same one operable for positive agonism namely, selective receptor state affinity. However, unlike agonists that have a selectively higher affinity for the receptor active state (to induce G-protein activation and subsequent physiological response) inverse agonists have a selectively higher affinity for the inactive receptor state and thus uncouple already spontaneously coupled [RaG] species in the system. 

One target type for which the molecular mechanism of efficacy has been partly elucidated is the G-protein-coupled receptor (GPCR). It is known that activation of GPCRs leads to an interaction of the receptor with separate membrane G-proteins to cause dissociation of the G-protein subunits and subsequent activation of effectors (see Chapter 2). For the purposes of binding, this process can lead to an aberration in the binding reaction as perceived in experimental binding studies. Specifically, the activation of the receptor with subsequent binding of that... 

Stimulation and inhibition of the enzyme by the GPCR-G-protein cycle occur by analogous mechanisms. Agonists induce hormone receptors to increase a Ga-GDP-GTP exchange and subsequent Ga 3y dissociation (GDP-a py + GTP GTP-ax + [3y + GDP) (Fig. 4). Consequently, agents that affect either the dissociation of either G or Gs, or the association of their respective as, a , or (3y subunits with adenylyl cyclase could affect rates of cAMP formation in enzyme preparations or in intact cells and tissues. There are several important examples. Gas is stably activated by poorly hydrolyzable analogs of GTP, e.g. GTPyS... 

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